Anti-PCSK9 antibodies and use thereof

ABSTRACT

The present invention is directed to antibodies and fragments thereof having binding specificity for PCSK9. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V H , V L  and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-PCSK9 antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-PCSK9 antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-PCSK9 antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with PCSK9.

RELATED APPLICATION DISCLOSURE

This application claims the benefit of U.S. Provisional Application Ser.No. 61/654,481, filed Jun. 1, 2012 and U.S. Provisional Application Ser.No. 61/644,065, filed May 8, 2012, each of which is hereby incorporatedby reference in its entirety.

The instant application contains a Sequence Listing which has beensubmitted in ASCII format via EFS-Web and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Mar. 10, 2013, isnamed 67858o791000.txt and is 621,210 bytes in size.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention pertains to novel antibodies and antibody fragments thatspecifically bind to human Proprotein Convertase Subtilisin-like/Kexintype 9 (hereinafter “PCSK9”) and compositions containing. In additionthe invention relates to nucleic acids encoding said antibodies andantibody fragments and the use thereof to express said antibodies andantibody fragments in desired host cells. Also, the invention relates totherapeutic and diagnostic use of these antibodies and antibodyfragments.

More particularly, the invention provides rabbit antibodies andhumanized and chimeric antibodies derived therefrom specific to PCSK9 aswell as antibody fragments specific to PCSK9 which include e.g., Fab′,F(ab′)₂, Fv, scFv fragments, SMIPs (small moleculeimmunopharmaceuticals), camelbodies, nanobodies, monovalent antibodiessuch as MetMab like antibodies, and IgNAR.

Further, the invention provides nucleic acids and host cells containingthat encode for and result in the expression of the subject anti-PCSK9antibodies, i.e., rabbit antibodies and antibody fragments and modifiedforms thereof including by way of example humanized and chimericantibodies derived therefrom as well as antibody fragments which includee.g., Fab′, F(ab′)₂, Fv, scFv fragments, SMIPs (small moleculeimmunopharmaceuticals), camelbodies, nanobodies, monovalent antibodiessuch as MetMab like antibodies, and IgNAR.

Also the invention relates to expression systems for the manufacture ofthe inventive anti-PCSK9 antibodies, including yeast, fungi, mammalian,and other cells useful for the manufacture of antibodies and antibodyfragments.

Also, the invention relates to novel antibodies and antibody fragmentsthat specifically bind to human PCSK9 which compete with and/orspecifically bind to the same or overlapping epitope(s) on PCSK9 as anyof the anti-PCSK9 antibodies and antibody fragments exemplified herein.

The invention further pertains to the in vivo use of the subjectanti-PCSK9 antibodies and antibody fragments alone or in associationwith other active agents or drugs. for blocking, inhibiting orneutralizing PCSK9.

The invention further pertains to the in vivo use of the subjectanti-PCSK9 antibodies and antibody fragments alone or in associationwith other active agents or drugs. for blocking or inhibiting theinteraction of PCSK9 with LDLR.

The invention also specifically relates to methods for treating orpreventing disorders of cholesterol or lipid homeostasis and disordersassociated therewith including by way of example hypercholesterolemia,hyperlipidemia, hypertriglyceridaemia, sitosterolemia, atherosclerosis,arteriosclerosis, coronary heart disease, metabolic syndrome, acutecoronary syndrome, xanthoma, hypertension, angina, obesity, diabetes andvascular inflammation, by the administration of the subject anti-PCSK9antibodies and antibody fragments, wherein the subject antibodies andantibody fragments may be used alone or in association with other activeagents.

The invention further specifically relates to methods of preventing ortreating diseases and disorders associated with PCSK9, e.g., diseasesassociated with increased or decreased levels of PCSK9 and/or mutationsin the PCSK9 gene that affect PCSK9 protein expression, primary sequenceand/or function by administering said antibodies or fragments thereofalone or in combination with other active agents.

The present invention further provides methods for improving bloodcholesterol markers associated with increased risk of heart diseaseusing the subject antibodies and antibody fragments alone or inassociation with other active agents. These markers include, but are notlimited to, high total cholesterol, high LDL, high total cholesterol toHDL ratio and high LDL-C to HDL ratio.

The present invention further provides methods for treating orpreventing any of the following conditions or complications associatedtherewith such as hypercholesterolemia, heart disease, metabolicsyndrome, diabetes, coronary heart disease, stroke, cardiovasculardiseases, Alzheimer's disease and generally dyslipidemias, which can bemanifested, for example, by an elevated total serum cholesterol,elevated LDL, elevated triglycerides, elevated VLDL, and/or low HDL.Some non-limiting examples of primary and secondary dyslipidemias thatcan be treated using an antibody or antibody fragment according to theinvention, either alone, or in combination with one or more other agentsinclude the metabolic syndrome, diabetes mellitus, familial combinedhyperlipidemia, familial hypertriglyceridemia, familialhypercholesterolemias, including heterozygous hypercholesterolemia,homozygous hypercholesterolemia, familial defective apoplipoproteinB-100; polygenic hypercholesterolemia; remnant removal disease, hepaticlipase deficiency; dyslipidemia secondary to any of the following:dietary indiscretion, hypothyroidism, drugs including estrogen andprogestin therapy, beta-blockers, and thiazide diuretics; nephroticsyndrome, chronic renal failure, Cushing's syndrome, primary biliarycirrhosis, glycogen storage diseases, hepatoma, cholestasis, acromegaly,insulinoma, isolated growth hormone deficiency, and alcohol-inducedhypertriglyceridemia.

In addition, antibody or antibody fragment according to the inventioncan also be useful in preventing or treating atherosclerotic diseases,such as, for example, coronary heart disease, coronary artery disease,peripheral arterial disease, stroke (ischaemic and hemorrhagic), anginapectoris, or cerebrovascular disease and acute coronary syndrome,myocardial infarction. In some embodiments, the antibody or antibodyfragment according to the invention is useful in reducing the risk of:nonfatal heart attacks, fatal and non-fatal strokes, certain types ofheart surgery, hospitalization for heart failure, chest pain in patientswith heart disease, and/or cardiovascular events because of establishedheart disease such as prior heart attack, prior heart surgery, and/orchest pain with evidence of clogged arteries. In some embodiments, theantibody or antibody fragment according to the invention and methods canbe used to reduce the risk of recurrent cardiovascular events.

The invention also particularly relates to the use of the subjectanti-PCSK9 antibodies and antibody fragments in any of theaforementioned therapeutic indications or conditions in combination withother drugs that are typically used to treat such disorders, wherein theantibody and other drug or agent may be co-administered or separatelyadministered. Non limiting examples of drugs that may be co-administeredwith the subject antibodies or antibody fragments or used in the sametherapeutic regimen include by way of example statins, ACE inhibitors,Angiotensin II receptor blockers (ARBs), Antiarrhythmics, AntiplateletDrugs, aspirin, beta blockers, amiodarone, digoxin, aspirin,anti-clotting agents, digoxin, diuretics, heart failure drugs,vasodilators, blood thinners, other anti-cholesterol drugs such asholestyramine (Questran), gemfibrozil (Lopid, Gemcor), Omacor, andpantethine, other anti-hypertensives, antidiabetigenic drugs such asAlpha-glucosidase inhibitors, Biguanides, Dipeptidyl peptidase-4inhibitors, Insulin therapies, Meglitinides, Sulfonylurea, andThiazolidinediones, and other drugs used to treat conditions wherein thetreated individual may have high cholesterol.

The invention further relates to compositions containing the subjectanti-PCSK9 antibodies or antibody fragments, especially compositions aresuitable for in vivo administration, e.g., subcutaneous, intravenous,intradermal, intranasal, rectal, vaginal, intrathecal, oral, and otheradministrable dosage forms.

The invention further relates to compositions containing the subjectanti-PCSK9 antibodies or antibody fragments, especially compositionssuitable for in vivo administration, e.g., subcutaneous, intravenous,intradermal, intranasal, rectal, vaginal, intrathecal, oral, and otheradministrable dosage forms which contain another active agent such asstatins, ACE inhibitors, Angiotensin II receptor blockers (ARBs),Antiarrhythmics, Antiplatelet Drugs, aspirin, beta blockers, amiodarone,digoxin, aspirin, anti-clotting agents, digoxin, diuretics, heartfailure drugs, vasodilators, blood thinners, other anti-cholesteroldrugs such as holestyramine (Questran), gemfibrozil (Lopid, Gemcor),Omacor, and pantethine, other anti-hypertensives, antidiabetigenic drugssuch as Alpha-glucosidase inhibitors, Biguanides, Dipeptidyl peptidase-4inhibitors, Insulin therapies, Meglitinides, Sulfonylurea, andThiazolidinediones, and other drugs used to treat conditions wherein thetreated individual may have high or aberrant lipid or cholesterollevels.

The invention further relates to storage stable forms containing thesubject anti-PCSK9 antibodies or antibody fragments, e.g.,lyophilisates, suspensions, and buffered or temperature stablecompositions.

The invention also pertains to methods of using the subject antibodiesand antibody fragments that specifically bind to PCSK9 in screeningassays to detect and monitor the levels of PCSK9 in serum samples,potentially for the diagnosis of diseases and disorders associated withPCSK9 or identifying individuals wherein the administration of thesubject antibodies and antibody fragments that specifically bind toPCSK9 may have a therapeutic or prophylactic effect or for monitoringthe effects of a treatment designed to modulate or neutralize PCSK9 orblock cholesterol synthesis, e.g., a treatment involving administrationof one of the subject anti-PCSK9 antibodies or antibody fragments.

2. Description of Related Art

Proprotein convertase subtilisin kexin type 9 (PCSK9) is a serineprotease involved in regulating the levels of the low densitylipoprotein receptor (LDLR) protein (Horton et al., 32(2) TrendsBiochem. Sci. 71-77 (2007); Seidah and Prat, 85(7) J. Mol. Med. 685-962007). In vitro experiments have shown that adding PCSK9 to HepG2 cellslowers the levels of cell surface LDLR (Benjannet et al., 279 J. Bio.Chem. 48865-75 (2004); Lagace et al., 116(11) J. Clin. Invest. 2995-3005(2006); Maxwell et al., 102(6) Proc. Nat. Acad. Sci. 2069-74 (2005);Park et al., 279 J. Biol. Chem. 50630-38 (2004)). (While this protein isgenerally referred to as PCSK9, it is noted that this protein andcorresponding gene has also been referred to in the patent andnon-patent literature by other names including PSEC0052, FH3, HCHOLA3,LDLCQ1, NARC-1, NARC1, PC9).

Experiments with mice have shown that increasing PCSK9 protein levelsdecreases levels of LDLR protein in the liver (Benjannet et al., 279 J.Bio. Chem. 48865-75 (2004); Lagace et al., 116(11) J. Clin. Invest.2995-3005 (2006); Maxwell et al., 102(6) Proc. Nat'l Acad. Sci. 2069-74(2005); Park et al., 279 J. Biol. Chem. 50630-38 (2004)), while PCSK9knockout mice have increased levels of LDLR in the liver (Rashid et al.,102(15) Proc. Nat'l Acad. Sci. 5374-79 (2005)). Additionally, varioushuman PCSK9 mutations that result in either increased or decreasedlevels of plasma LDL-C have been identified (Kotowski et al., 78(3) Am.J. Hum. Genet. 410-22 (2006); Zhao et al., 79(3) Am. J. Hum. Genet.514-23 (2006)). PCSK9 has been shown to directly interact with the LDLRprotein, be endocytosed along with the LDLR, and co-immunofluoresce withthe LDLR throughout the endosomal pathway (Lagace et al., 116(11) J.Clin. Invest. 2995-3005 (2006)). Several mutations in human PCSK9 causegain-of-function effects in humans, including hypercholesterolemia,increased LDL-C cholesterol levels, and increased risk of coronary heartdisease. (Garnier, 11(3) Am. J. Cardiovasc. Drugs 145-52 (2011)). Evenrarer human PCSK9 mutations induce loss-of-function, resulting inlowered LDL-C levels and a 88% reduction in coronary heart disease risk.(Id.) PCSK9 interacts with the LDLR via the EGF domain and the complexis internalized. In the endosome, the lower pH results in an increasedaffinity between PCSK9 and LDLR and the complex is targeted fordegradation in the lysosome (Horton et al., April Supp., J. Lipid Res.S172-177 (2009), Sci. 928-33 (2003)).

Several lines of evidence demonstrate that PCSK9, in particular, lowersthe amount of hepatic LDLR protein and thus compromises the liver'sability to remove LDL cholesterol from the circulation.Adenovirus-mediated overexpression of PCSK9 in the livers of miceresults in the accumulation of circulating LDL-C due to a dramatic lossof hepatic LDLR protein, with no effect on LDLR mRNA levels; Benjannetet al., 2004 J. Biol. Chem. 279:48865-48875; Maxwell & Breslow, 2004PNAS 101:7100-7105; Park et al., 2004 J. Biol. Chem. 279:50630-50638;and Lalanne et al., 2005 J. Lipid Res. 46:1312-1319. The effect of PCSK9overexpression on raising circulating LDL-C levels in mice is completelydependent on the expression of LDLR, again, indicating that theregulation of LDL-C by PCSK9 is mediated through downregulation of LDLRprotein. In agreement with these findings, mice lacking PCSK9 or inwhich PCSK9 mRNA has been lowered by antisense oligonucleotideinhibitors have higher levels of hepatic LDLR protein and a greaterability to clear circulating LDL-C; Rashid et al., 2005 PNAS102:5374-5379; and Graham et al., 2007 J. Lipid Res. 48(4):763-767. Inaddition, lowering PCSK9 levels in cultured human hepatocytes by siRNAalso results in higher LDLR protein levels and an increased ability totake up LDL-C; Benjannet et al., 2004 J. Biol. Chem. 279:48865-48875;and Lalanne et al., 2005 J Lipid Res. 46:1312-1319. Together, these dataindicate that PCSK9 action leads to increased LDL-C by lowering LDLRprotein levels.

A number of mutations in the gene PCSK9 have also been conclusivelyassociated with autosomal dominant hypercholesterolemia (“ADH”), aninherited metabolism disorder characterized by marked elevations of lowdensity lipoprotein (“LDL”) particles in the plasma which can lead topremature cardiovascular failure; see Abifadel et al., 2003 NatureGenetics 34:154-156; Timms et al., 2004 Hum. Genet. 114:349-353; Leren,2004 Clin. Genet. 65:419-422. A later-published study on the S127Rmutation of Abifadel et al., supra, reported that patients carrying sucha mutation exhibited higher total cholesterol and apoB100 in the plasmaattributed to (1) an overproduction of apoB100-containing lipoproteins,such as low density lipoprotein (“LDL”), very low density lipoprotein(“VLDL”) and intermediate density lipoprotein (“IDL”), and (2) anassociated reduction in clearance or conversion of said lipoproteins;Ouguerram et al., 2004 Arterioscler. Thromb. Vasc. Biol. 24:1448-1453.

PCSK9 is a prohormone-proprotein convertase in the subtilisin (S8)family of serine proteases (Seidah et al., 100(3) Proc. Nat'l Acad. Sci.928-33 (2003)). Humans have nine prohormone-proprotein convertases thatcan be divided between the S8A and S8B subfamilies (Rawlings et al., 34Nucleic Acids Research D270-72 (2006)). Furin, PC1/PC3, PC2, PACE4, PC4,PC5/PC6 and PC7/PC8/LPC/SPC7 are classified in subfamily S8B. Crystaland NMR structures of different domains from mouse furin and PC1 revealsubtilisin-like pro- and catalytic domains, and a P domain directlyC-terminal to the catalytic domain (Henrich et al., 10(7) NatureStructural Bio. 520-26 (2005); Tangrea et al., 320(4) J. Mol. Bio.801-12 (2002)). Based on the amino acid sequence similarity within thissubfamily, all seven members are predicted to have similar structures(Henrich et al., 345(2) J. Mol. Bio. 211-27 (2005)). SKI-1/S1P and PCSK9are classified in subfamily S8A. Sequence comparisons with theseproteins also suggest the presence of subtilisin-like pro- and catalyticdomains (Sakai et al., 2(4) Mol. Cell. 505-15 (1998); Seidah et al.,100(3) Proc. Nat'l Acad. Sci. 928-33 (2003); Seidah et al., 96(4) Proc.Nat'l Acad. Sci. 1321-26 (1999)). In these proteins, the amino acidsequence C-terminal to the catalytic domain is more variable and doesnot suggest the presence of a P domain.

Prohormone-proprotein convertases are expressed as zymogens, and theymature through a multi step process. The function of the pro-domain inthis process is two-fold. The pro-domain first acts as a chaperone andis required for proper folding of the catalytic domain (Ikemura et al.,262 J. Biol. Chem. 7859-64 (1987)). Once the catalytic domain is folded,autocatalysis occurs between the pro-domain and catalytic domain.Following this initial cleavage reaction, the pro-domain remains boundto the catalytic domain where it then acts as an inhibitor of catalyticactivity (Fu et al., 275 J. Biol. Chem. 16871-78 (2000)). Whenconditions are correct, maturation proceeds with a second autocatalyticevent at a site within the pro-domain (Anderson et al., 16 Nature1508-18 (1997)). After this second cleavage event occurs the pro-domainand catalytic domain dissociate, giving rise to an active protease.

Autocatalysis of the PCSK9 zymogen occurs between Gln152 and Ser153(VFAQ|SIP) (Naureckiene et al., 420(1) Archives of Biochem. & Biophysics55-67 (2003)), and has been shown to be required for its secretion fromcells (Seidah et al., 100(3) Proc. Nat'l Acad. Sci. 928-33 (2003)). Asecond autocatalytic event at a site within PCSK9's pro-domain has notbeen observed. Purified PCSK9 is made up of two species that can beseparated by non-reducing SDS-PAGE; the pro-domain at 17 Kd, and thecatalytic plus C-terminal domains at 65 Kd. PCSK9 has not been isolatedwithout its inhibitory pro-domain, and measurements of PCSK9's catalyticactivity have been variable (Naureckiene et al., 420(1) Archives ofBiochem. & Biophysics 55-67 (2003); Seidah et al., 100(3) Proc. Nat'lAcad. Sci. 928-33 (2003)).

Accordingly, there is substantial evidence indicating that PCSK9 plays arole in the regulation of LDL; that the expression or upregulation ofPCSK9 is associated with increased plasma levels of LDL cholesterol,that the corresponding inhibition or lack of expression of PCSK9 isassociated with reduced LDL cholesterol plasma levels; and thatdecreased levels of LDL cholesterol are associated with sequencevariations in PCSK9 have been found to confer protection againstcoronary heart disease; Cohen, 2006 N. Engl. J. Med. 354:1264-1272.

In clinical trials, reductions in LDL cholesterol levels have beendirectly related to the rate of coronary events; Law et al., 2003 BMJ326:1423-1427. Also, moderate lifelong reduction in plasma LDLcholesterol levels was found to correlate with a substantial reductionin the incidence of coronary events; Cohen et al., supra. This was thecase even in populations with a high prevalence of non-lipid-relatedcardiovascular risk factors; supra. Accordingly, there is great benefitto be reaped from the managed control of LDL cholesterol levels.

Based thereon, the identification of other molecules which may be usedto modulate cholesterol levels and block or inhibit or neutralize theactivity of PCSK9 would be of great interest. The present inventionadvances these interests by providing novel antagonists of PCSK9 for usefor in blocking, inhibiting or neutralizing one or more of theactivities of PCSK9 and/or in blocking the interaction of PCSK9 withLDLR and/or for the treatment of therapeutic conditions identifiedherein especially those involving or associated with high or aberrantlipid or cholesterol levels.

BRIEF SUMMARY OF THE INVENTION

The present invention provides novel antibodies and antibody fragmentsthat specifically bind PCSK9, as well as antibodies that compete withsuch antibodies or antibody fragments, or which specifically bind to thesame or overlapping epitope, or antibodies which contain any or all ofthe CDRs of the novel antibodies and antibody fragments exemplifiedherein. These antibodies and antibody fragments may be used to block,inhibit or neutralize the in vivo effects of serum PCSK9 in vivo, and insome embodiments specifically inhibit or block the PCSK9/LDLR bindinginteraction and thereby inhibit or neutralize one or all of thebiological effects associated with this binding interaction.

Particularly, the invention provides novel antibodies and antibodyfragments that specifically bind to human PCSK9 and compositionscontaining. More particularly, the invention provides rabbit anti-PCSK9antibodies and antibody fragments as well as humanized and chimericantibodies and antibody fragments derived therefrom such as Fab, Fab′,F(ab′)₂, Fv, scFv fragments, SMIPs (small moleculeimmunopharmaceuticals), camelbodies, nanobodies, monovalent antibodiessuch as MetMab like antibodies, and IgNAR.

The invention further describes the use of the subject anti-PCSK9antibodies and antibody fragments for treating any subject whereinblocking, inhibiting or neutralizing the in vivo effect of PCSK9 orblocking or inhibiting the interaction of PCSK9 and LDLR istherapeutically desirable, wherein the subject anti-PCSK9 antibodies orantibody fragments may be used alone or in association with other activeagents or drugs.

The invention also describes methods for treating or preventingdisorders of cholesterol or lipid homeostasis and disorders which may beassociated therewith including by way of example hypercholesterolemia,hyperlipidemia, hypertriglyceridaemia, sitosterolemia, atherosclerosis,arteriosclerosis, coronary heart disease, metabolic syndrome, acutecoronary syndrome, vascular inflammation, diabetes, obesity, angina,hypertension and xanthoma by the administration of the subjectanti-PCSK9 antibodies and antibody fragments that specifically bind toPCSK9, wherein the subject antibodies and antibody fragments may be usedalone or in association with other active agents.

The invention further provides methods of preventing or treatingdiseases and disorders associated with PCSK9, e.g., diseases associatedwith increased or decreased levels of PCSK9 and/or mutations in thePCSK9 gene that affect PCSK9 protein expression, primary sequence and/orfunction by administering said antibodies or fragments thereof alone orin combination with other active agents.

The invention broadly encompasses the use of the subject anti-PCSK9antibodies and fragments in treating any subject having a condition orat risk of developing a condition wherein modulation of lipid orcholesterol levels is clinically desirable or where the subject has acondition that is often associated with high lipids or cholesterol.

Also specifically the present invention provides methods for treating orpreventing disorders of cholesterol or lipid homeostasis and disordersand complications associated therewith, e.g., hypercholesterolemia,hyperlipidemia, hypertriglyceridaemia, sitosterolemia, atherosclerosis,arteriosclerosis, coronary heart disease, metabolic syndrome, acutecoronary syndrome, vascular inflammation, xanthoma, hypertension, anginaand related conditions by administration of the subject anti-PCSK9antibodies and antibody fragments alone or in association with otheractive agents.

The present invention further provides methods for improving bloodcholesterol markers associated with increased risk of heart diseaseusing the subject antibodies and antibody fragments alone or inassociation with other active agents. These markers include, but are notlimited to, high total cholesterol, high LDL, high total cholesterol toHDL ratio and high LDL-C to HDL ratio.

The invention also particularly relates to the use of the subjectanti-PCSK9 antibodies and antibody fragments in any of theaforementioned therapeutic indications or conditions in combination withother drugs that are typically used to treat such disorders, wherein theantibody and other drug or agent may be co-administered or separatelyadministered. Examples of drugs include that may be co-administered withthe subject anti-PCSK9 antibodies or antibody fragments or in the sametherapeutic regimen include by way of example statins, ACE inhibitors,Angiotensin II receptor blockers (ARBs), Antiarrhythmics, AntiplateletDrugs, aspirin, beta blockers, amiodarone, digoxin, aspirin,anti-clotting agents, digoxin, diuretics, heart failure drugs,vasodilators, blood thinners, other anti-cholesterol drugs such asholestyramine (Questran), gemfibrozil (Lopid, Gemcor), Omacor, andpantethine, other anti-hypertensives, antidiabetigenic drugs such asAlpha-glucosidase inhibitors, Biguanides, Dipeptidyl peptidase-4inhibitors, Insulin therapies, Meglitinides, Sulfonylurea, andThiazolidinediones, and other drugs used to treat conditions wherein thetreated individual may have high cholesterol or aberrant lipid levels orlipid metabolism.

The invention further relates to compositions containing the subjectanti-PCSK9 antibodies or antibody fragments, especially compositions aresuitable for in vivo administration, e.g., subcutaneous, intravenous,intradermal, intranasal, intrathecal, vaginal, rectal, and otherinjectable or topical administrable dosage forms.

The present invention further provides methods for treating orpreventing any of the following conditions or complications associatedtherewith such as hypercholesterolemia, heart disease, metabolicsyndrome, diabetes, coronary heart disease, stroke, cardiovasculardiseases, Alzheimer's disease and generally dyslipidemias, which can bemanifested, for example, by an elevated total serum cholesterol,elevated LDL, elevated triglycerides, elevated VLDL, and/or low HDL.Some non-limiting examples of primary and secondary dyslipidemias thatcan be treated using an antibody or antibody fragment according to theinvention, either alone, or in combination with one or more other agentsinclude the metabolic syndrome, diabetes mellitus, familial combinedhyperlipidemia, familial hypertriglyceridemia, familialhypercholesterolemias, including heterozygous hypercholesterolemia,homozygous hypercholesterolemia, familial defective apoplipoproteinB-100; polygenic hypercholesterolemia; remnant removal disease, hepaticlipase deficiency; dyslipidemia secondary to any of the following:dietary indiscretion, hypothyroidism, drugs including estrogen andprogestin therapy, beta-blockers, and thiazide diuretics; nephroticsyndrome, chronic renal failure, Cushing's syndrome, primary biliarycirrhosis, glycogen storage diseases, hepatoma, cholestasis, acromegaly,insulinoma, isolated growth hormone deficiency, and alcohol-inducedhypertriglyceridemia.

In addition, the present invention further provides methods for use ofthe subject anti-PCSK9 antibody or antibody fragment in preventing ortreating atherosclerotic diseases, such as, for example, coronary heartdisease, coronary artery disease, peripheral arterial disease, stroke(ischaemic and hemorrhagic), angina pectoris, or cerebrovascular diseaseand acute coronary syndrome, myocardial infarction; in reducing the riskof: nonfatal heart attacks, fatal and non-fatal strokes, certain typesof heart surgery, hospitalization for heart failure, chest pain inpatients with heart disease, and/or cardiovascular events because ofestablished heart disease such as prior heart attack, prior heartsurgery, and/or chest pain with evidence of clogged arteries; and toreduce the risk of recurrent cardiovascular events.

More specifically, the invention provides compositions containing atleast one of the subject anti-PCSK9 antibodies or antibody fragments,especially compositions which are suitable for in vivo administration,e.g., subcutaneous, intravenous, intradermal, intranasal, intrathecal,vaginal, rectal, oral and other injectable or topical dosage forms whichoptionally may contain another active agent such as statins, ACEinhibitors, Angiotensin II receptor blockers (ARBs), Antiarrhythmics,Antiplatelet Drugs, aspirin, beta blockers, amiodarone, digoxin,aspirin, anti-clotting agents, digoxin, diuretics, heart failure drugs,vasodilators, blood thinners, other anti-cholesterol drugs such asholestyramine (Questran), gemfibrozil (Lopid, Gemcor), Omacor, andpantethine, other anti-hypertensives, antidiabetigenic drugs such asAlpha-glucosidase inhibitors, Biguanides, Dipeptidyl peptidase-4inhibitors, Insulin therapies, Meglitinides, Sulfonylurea, andThiazolidinediones, and other drugs used to treat conditions wherein thetreated individual may have high cholesterol. The invention alsoprovides novel dosage regimens using the subject anti-PCSK9 antibodiesor antibody fragments, alone or in association with another activeagent, especially subcutaneous, oral and intravenous dosing regimens.

The invention further relates to storage stable forms containing thesubject anti-PCSK9 antibodies or antibody fragments, e.g.,lyophilisates, suspensions, and buffered or temperature stablecompositions containing.

The invention also provides methods of using the subject antibodies andantibody fragments that specifically bind to PCSK9 in screening assaysto detect and monitor the levels of PCSK9 in serum samples, potentiallyfor the diagnosis of diseases and disorders associated with PCSK9 or inidentifying individuals wherein the administration of the subjectantibodies and antibody fragments that specifically bind to PCSK9 mayhave a therapeutic or prophylactic effect or for monitoring the effectsof a treatment designed to modulate or neutralize PCSK9 or blockcholesterol synthesis, e.g., a treatment involving administration of oneof the subject or other anti-PCSK9 antibodies or antibody fragments.

Further, the invention provides specific anti-PCSK9 antibodies andfragments thereof and diagnostic or pharmaceutical compositionscontaining having particular epitopic specificity for PCSK9, andpreferably antibodies or fragments thereof having high affinity oravidity and/or other desired functional properties.

In addition, the invention relates to any therapeutic or diagnostic useof the antibodies described herein, or antibodies competing therewith orpossessing the same or overlapping epitopic specificity, preferablyantibodies or antibody fragments comprising one or all of the CDRs ofone of the exemplified anti-PCSK9 antibodies or antibody fragments, ormore preferably an antibody comprising one or more variable or CDRsequences which possess at least 80, 90, or 95, 96, 97, 98, 99 or 100%identity to any of the VH, VL and CDR polypeptides described herein, andto polynucleotides encoding these antibodies and antibody fragments andhost cells containing. A preferred embodiment of the invention isdirected to chimeric or humanized antibodies and fragments thereof (suchas Fab or Fv or monovalent fragments) capable of binding to PCSK9, andpreferably which inhibit, block or neutralize the biological activitiesof PCSK9 or which block or inhibit the binding of PCSK9 to LDLR.

In some aspects, the invention comprises an isolated antibody orantibody fragment that competes for binding to PCSK9 or binds with thesame or an overlapping epitope on PCSK9 as an antibody or antibodyfragment according to the invention.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a patient, comprising administering to a patient in need thereof aneffective amount of at least one anti-PCSK9 antibody or antibodyfragment according to the invention.

In some aspects, the invention comprises a method of inhibiting bindingof PCSK9 to LDLR in a subject in need thereof comprising administeringan effective amount of at least one anti-PCSK9 antibody or antibodyfragment according to the invention.

In some aspects, the invention comprises an anti-PCSK9 antibody orantibody fragment according to the invention that binds to PCSK9 with aKD that is less than 100 nM.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, the method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orantibody fragment according to the invention simultaneously orsequentially with another active agent, e.g., one that reducescholesterol levels or which elevates the availability of LDLR protein.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject, the method comprising administering toa subject an effective amount of at least at least one anti-PCSK9antibody or antibody fragment according to the invention.

In some aspects, the invention comprises a method of lowering serumcholesterol level in a subject, the method comprising administering to asubject an effective amount of at least one anti-PCSK9 antibody orantibody fragment according to the invention, simultaneously orsequentially with another agent that elevates the availability of LDLRprotein or which reduces serum cholesterol.

In some aspects, the invention comprises a method of increasing LDLRprotein level in a subject, the method comprising administering to asubject an effective amount of at least one anti-PCSK9 antibody orantibody fragment according to the invention.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject, the method comprising administering to asubject an effective amount of at least one at least one anti-PCSK9antibody or antibody fragment according to the invention simultaneouslyor sequentially with an agent that elevates the availability of LDLRprotein or one which reduces serum cholesterol.

In some aspects, the invention further provides methods of preventing ortreating diseases and disorders associated with PCSK9, e.g., diseasesassociated with increased or decreased levels of PCSK9 and/or mutationsin the PCSK9 gene that affect PCSK9 protein expression, primary sequenceand/or function by administering at least one at least one anti-PCSK9antibody or antibody fragment according to the invention in combinationwith other agents.

In other aspects the present invention provides methods for treating orpreventing disorders of cholesterol or lipid homeostasis and disordersand complications associated therewith, e.g., hypercholesterolemia,hyperlipidemia, hypertriglyceridaemia, sitosterolemia, atherosclerosis,arteriosclerosis, coronary heart disease, metabolic syndrome, acutecoronary syndrome, vascular inflammation, xanthoma and relatedconditions using the subject anti-PCSK9 antibodies and antibodyfragments.

In other specific aspects the present invention provides methods fortreating or preventing disorders of cholesterol or lipid homeostasis anddisorders and complications associated therewith, e.g.,hypercholesterolemia, hyperlipidemia, hypertriglyceridaemia,sitosterolemia, atherosclerosis, arteriosclerosis, coronary heartdisease, metabolic syndrome, acute coronary syndrome, vascularinflammation, xanthoma and other conditions such as obesity,hypertension, diabetes, wherein the subject is treated with the subjectanti-PCSK9 antibodies and antibody fragments, in combination with otherdrugs used to treat such disorders such as e.g., statins, ACEinhibitors, Angiotensin II receptor blockers (ARBs), Antiarrhythmics,Antiplatelet Drugs, aspirin, beta blockers, amiodarone, digoxin,aspirin, anti-clotting agents, digoxin, diuretics, heart failure drugs,vasodilators, blood thinners, other anti-cholesterol drugs such asholestyramine (Questran), gemfibrozil (Lopid, Gemcor), Omacor, andpantethine, other anti-hypertensives, antidiabetigenic drugs such asAlpha-glucosidase inhibitors, Biguanides, Dipeptidyl peptidase-4inhibitors, Insulin therapies, Meglitinides, Sulfonylurea, andThiazolidinediones, and other drugs used to treat conditions wherein thetreated individual may have high cholesterol.

ACE inhibitors may be used in combination with the subject anti-PCSK9antibodies and antibody fragments wherein the moieties may be jointly orseparately administered by the same or different means of administrationinclude by way of example: Capoten (captopril), Vasotec (enalapril),Prinivil, Zestril (lisinopril), Lotensin (benazepril), Monopril(fosinopril), Altace (ramipril), Accupril (quinapril), Aceon(perindopril), Mavik (trandolapril), Univasc (moexipril),

ARBs may be used in combination with the subject anti-PCSK9 antibodiesand antibody fragments wherein the moieties may be jointly or separatelyadministered by the same or different means of administration include byway of example: Cozaar (losartan), Diovan (valsartan), Avapro(irbesartan), Atacand (candesartan), and Micardis (telmisartan).

Antiarrhythmics may be used in combination with the subject anti-PCSK9antibodies and antibody fragments include by way of example: Tambocor(flecamide), Procanbid (procainamide), Cordarone (amiodarone), andBetapace (sotalol).

Anticlotting agents which may be used in combination with the subjectanti-PCSK9 antibodies and antibody fragments wherein the moieties may bejointly or separately administered by the same or different means ofadministration include: Tissue plasminogen activator (TPA),Tenecteplase, Alteplase, Urokinase, Reteplase, and Streptokinase.

Beta-blockers may be used in combination with the subject anti-PCSK9antibodies and antibody fragments wherein the moities may be jointly orseparately administered by the same or different means of administrationinclude by way of example: Sectral (acebutolol), Zebeta (bisoprolol),Brevibloc (esmolol), Inderal (propranolol), Tenormin (atenolol),Normodyne, Trandate (labetalol), Coreg (carvedilol), Lopressor, andToprol-XL (metoprolol).

Calcium channel blockers which may be used in combination with thesubject anti-PCSK9 antibodies and antibody fragments wherein the moitiesmay be jointly or separately administered by the same or different meansof administration include by way of example: Norvasc (amlodipine),Plendil (felodipine), Cardizem, Cardizem CD, Cardizem SR, Dilacor XR,Diltia XT, Tiazac (diltiazem), Calan, Calan SR, Covera-HS, Isoptin,Isoptin SR, Verelan, Verelan PM (verapamil), Adalat, Adalat CC,Procardia, Procardia XL (nifedipine), Cardene, Cardene SR (nicardipine),Sular (nisoldipine), Vascor (bepridil), and Caduet which is acombination of a statin cholesterol drug and amlodipine.

Diuretics which may be used in combination with the subject anti-PCSK9antibodies and antibody fragments wherein the moities may be jointly orseparately administered by the same or different means of administrationinclude by way of example Lasix (furosemide), Bumex (bumetanide),Demadex (torsemide), Esidrix (hydrochlorothiazide), Zaroxolyn(metolazone), and Aldactone (spironolactone).

Heart failure drugs which may be used in combination with the subjectanti-PCSK9 antibodies and antibody fragments wherein the moities may bejointly or separately administered by the same or different means ofadministration include by way of example Dobutrex (dobutamine), andPrimacor (milrinone).

Vasodilators which may be used in combination with the subjectanti-PCSK9 antibodies and antibody fragments wherein the moities may bejointly or separately administered by the same or different means ofadministration include by way of example Dilatrate-SR, Iso-Bid, Isonate,Isorbid, Isordil, Isotrate, Sorbitrate (isosorbide dinitrate), IMDUR(isorbide mononitrate), and BiDil (hydralazine with isosorbidedinitrate.

Blood thinners which may be used in combination with the subjectanti-PCSK9 antibodies and antibody fragments wherein the moities may bejointly or separately administered by the same or different means ofadministration include by way of example Warfarin (coumadin), Heparin,Lovenox, and Fragmin.

In other aspects the present invention further provides methods forimproving blood cholesterol markers associated with increased risk ofheart disease using the subject antibodies and antibody fragments inassociation with any of the foregoing or other actives wherein themoities may be jointly or separately administered by the same ordifferent means of administration. These markers include, but are notlimited to, high total cholesterol, high LDL-C, high total cholesterolto HDL ratio and high LDL-C to HDL ratio.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody protein or fragment according to the inventionand another active, e.g., one of the actives above-identified, e.g., anagent that elevates the availability of LDLR protein levels or an agentwhich blocks or inhibits cholesterol synthesis. In some embodiments, theagent that blocks cholesterol synthesis comprises a statin. The statinin some instances potentially may further elevate LDLR levels. In someembodiments, the statin is selected from the group consisting ofatorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin,pitavastatin, pravastatin, rosuvastatin, simvastatin, and somecombination thereof. In some embodiments, the statin is further combinedwith niacin, an absorption inhibitor (ezetimibe), a lipid modifyingagent, or a combination thereof. In some embodiments, the agent thatelevates the availability of LDLR protein levels comprises a cytokinesuch as oncostatin M, or a hormone like estrogen, and/or a herbal moietysuch as berberine.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody protein or fragment or variant thereof accordingto the invention and an agent that increases high density lipoprotein(HDL) and/or decreases triglyceride levels. In some embodiments, theagent that increases high density lipoprotein (HDL) and/or decreasestriglyceride levels comprises a fibrate. In some embodiments, thefibrate is selected from the group consisting of bezafibrate,ciprofibrate, clofibrate, gemfibrozil, fenofibrate, and some combinationthereof.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody protein or fragment according to the inventionand a bile acid sequestering agent. In some embodiments, the bilesequestering agent is selected from the group consisting ofcholestyramine, colesevelam, colestipol, and some combination thereof.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or antibody fragment according to the inventionand an agent that decreases cholesterol absorption in the intestine. Insome embodiments, the agent that decreases cholesterol absorption in theintestine is ezetimibe.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that decreases cholesterol levels. In some embodiments, the agentthat decreases cholesterol levels is selected from the group consistingof certain anti-psychotic agents, certain HIV protease inhibitors,dietary factors such as high fructose, sucrose, cholesterol or certainfatty acids and certain nuclear receptor agonists and antagonists forRXR, RAR, LXR, FXR.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and a PPARgamma agonist, PPAR alpha/gamma agonist, squalene synthase inhibitor,CETP inhibitor, anti-hypertensive, anti-diabetic agent (such assulphonyl ureas, insulin, GLP-1 analogs, DDPIV inhibitors), ApoBmodulator, MTP inhibitors, arteriosclerosis obliterans treatments, or acombination thereof.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that increases HDL levels. In some embodiments, the agent thatincreases HDL levels is niacin, also known as nicotinic acid. In someembodiments, the niacin is a slow-release formulation.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that inhibits hepatic triglyceride production and/or very lowdensity lipoprotein (VLDL) secretions. In some embodiments, the agentthat inhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions is acipimox.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that decreases lipid absorption in the intestine. In someembodiments, the agent that decreases lipid absorption in the intestineis selected from the group consisting of orlistat, lipstatin, and somecombination thereof.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that is an anti-hypertensive and/or treats angina. In someembodiments, the agent that is an anti-hypertensive and/or treats anginais amlodipine.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and atleast one other agent, wherein the combination allows for mitigation ofundesirable side-effects of at least one other agent. In someembodiments, the antibody or antibody fragment according to theinvention and the at least one other agent are administeredconcurrently. In some embodiments, the antibody or antibody fragmentaccording to the invention or fragment or variant thereof and at leastone other agent are not administered simultaneously, with the antibodyor antibody fragment according to the invention being administeredbefore or after the at least one other agent is administered.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and atleast one other agent for treating a condition associated with aberrantcholesterol or for treating a condition wherein the individuals oftenhave high cholesterol. For example, an antibody protein or fragment orvariant thereof according to the invention may be combined orco-administered with other drugs such as ACE inhibitors, Angiotensin IIreceptor blockers (ARBs), Antiarrhythmics, Antiplatelet Drugs, aspirin,beta blockers, amiodarone, digoxin, aspirin, anti-clotting agents,digoxin, diuretics, heart failure drugs, vasodilators, blood thinners,other anti-cholesterol drugs such as holestyramine (Questran),gemfibrozil (Lopid, Gemcor), Omacor, and pantethine, otheranti-hypertensives, antidiabetigenic drugs such as Alpha-glucosidaseinhibitors, Biguanides, Dipeptidyl peptidase-4 inhibitors, Insulintherapies, Meglitinides, Sulfonylurea, and Thiazolidinediones, or otherdrugs used to treat conditions wherein the treated individual may havehigh cholesterol. Examples of such drugs are identified supra. In someembodiments, the antibody or fragment according to the invention and atleast one other agent are not administered simultaneously, with theantibody or antibody fragment according to the invention beingadministered before or after the at least one other agent isadministered.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelin a patient comprising administering to a patient in need thereof aneffective amount of at least one isolated antibody or antibody fragmentaccording to the invention as disclosed herein. In some embodiments, thecondition is hypercholesterolemia.

In some aspects, anti-PCSK9 antibodies or fragments according to theinvention bind to PCSK9 with a KD that is less than about 100 nM. Insome embodiments, the anti-PCSK9 antibodies or fragments according tothe invention bind PCSK9 with a KD that is between about 10 and about100 nM. In some embodiments, the anti-PCSK9 antibodies or fragmentsaccording to the invention bind PCSK9 with a KD that is less than about10 nM. In some embodiments, the antibody or fragment that binds PCSK9has a KD that is between about 1 and about 10 nM. In some embodiments,the anti-PCSK9 antibodies or fragments according to the invention thatbinds PCSK9 has a KD that is less than about 1 nM. In some embodiments,the anti-PCSK9 antibodies or fragments according to the invention has aKD that is between about 0.1 and about 1 nM. In some embodiments, theanti-PCSK9 antibodies or fragments according to the invention that bindsPCSK9 has a KD that is between about 0.1 and about 0.5 nM. In someembodiments, the anti-PCSK9 antibodies or fragments according to theinvention binds PCSK9 with a KD that is between about 0.01 and about 0.1nM. In some embodiments, the anti-PCSK9 antibodies or fragmentsaccording to the invention binds PCSK9 with a KD that is between about0.1 and about 10 nM. In some embodiments, the anti-PCSK9 antibodies orfragments according to the invention bind PCSK9 with a KD that isbetween 0.120 and about 7.99 nM.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that decreases cholesterol and which optionally furtherelevates the availability of LDLR protein. In some embodiments, theagent that reduces cholesterol and which optionally elevates theavailability of LDLR protein comprises a statin. In some embodiments,the statin is selected from the group consisting of atorvastatin,cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin,pravastatin, rosuvastatin, simvastatin, and some combination thereof. Insome embodiments, the statin is combined with niacin, an absorptioninhibitor (ezetimibe), a lipid modifying agent, or a combinationthereof. In some embodiments, the agent that elevates the availabilityof LDLR protein levels comprises certain cytokines like oncostatin M,estrogen, and/or certain herbal ingredients such as berberine.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that increases high density lipoprotein (HDL) and/or decreasestriglyceride levels. In some embodiments, the agent that increases highdensity lipoprotein (HDL) and/or decreases triglyceride levels comprisesa fibrate. In some embodiments, the fibrate is selected from the groupconsisting of bezafibrate, ciprofibrate, clofibrate, gemfibrozil,fenofibrate, and some combination thereof.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially witha bile acid sequestering agent. In some embodiments, the bile acidsequestering agent is selected from the group consisting ofcholestyramine, colesevelam, colestipol, and some combination thereof.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated anti-PCSK9antibodies or fragments according to the invention simultaneously orsequentially with an agent that decreases cholesterol absorption in theintestine. In some embodiments, the agent that decreases cholesterolabsorption in the intestine is ezetimibe.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that decreases cholesterol levels. In some embodiments, theagent that decreases cholesterol levels is selected from the groupconsisting of certain anti-psychotic agents, certain HIV proteaseinhibitors, dietary factors such as high fructose, sucrose, cholesterolor certain fatty acids and certain nuclear receptor agonists andantagonists for RXR, RAR, LXR, FXR.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that increases HDL levels. In some embodiments, the agent thatincreases HDL levels is niacin, also known as nicotinic acid. In someembodiments, the niacin is a slow-release formulation.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that inhibits hepatic triglyceride production and/or very lowdensity lipoprotein (VLDL) secretions. In some embodiments, the agentthat inhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions is acipimox.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that decreases lipid absorption in the intestine. In someembodiments, the agent that decreases lipid absorption in the intestineis selected from the group consisting of orlistat, lipstatin, and somecombination thereof.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that is an anti-hypertensive and/or treats angina. In someembodiments, the agent that is an anti-hypertensive and/or treats anginais amlodipine.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withat least one other agent, wherein the combination allows for mitigationof undesirable side-effects of at least one other agent. In someembodiments, the anti-PCSK9 antibodies or fragments according to theinvention and the at least one other agent are administeredconcurrently. In some embodiments, the antibody or antibody fragmentaccording to the invention and at least one other agent are notadministered simultaneously, with the anti-PCSK9 antibodies or fragmentsaccording to the invention being administered before or after the atleast one other agent is administered.

In some aspects, the invention comprises a method of lowering the serumcholesterol level in a subject. The method comprises administering to asubject an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat reduces cholesterol and which optionally further optionallyelevates the availability of LDLR protein. In some embodiments, theagent that reduces cholesterol and which may further elevate theavailability of LDLR protein comprises a statin. In some embodiments,the statin is selected from the group consisting of atorvastatin,cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin,pravastatin, rosuvastatin, simvastatin, and some combination thereof. Insome embodiments, the statin is combined with niacin, an absorptioninhibitor (ezetimibe), a lipid modifying agent, or a combinationthereof. In some embodiments, the agent that elevates the availabilityof LDLR protein levels comprises certain cytokines like oncostatin M,estrogen, and/or certain herbal ingredients such as berberine.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat increases high density lipoprotein (HDL) and/or decreasestriglyceride levels. In some embodiments, the agent that increases highdensity lipoprotein (HDL) and/or decreases triglyceride levels comprisesa fibrate. In some embodiments, the fibrate is selected from the groupconsisting of bezafibrate, ciprofibrate, clofibrate, gemfibrozil,fenofibrate, and some combination thereof.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with a bileacid sequestering agent. In some embodiments, the bile acid sequesteringagent is selected from the group consisting of cholestyramine,colesevelam, colestipol, and some combination thereof.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one at least one anti-PCSK9 antibody orfragment according to the invention, simultaneously or sequentially withan agent that decreases cholesterol absorption in the intestine. In someembodiments, the agent that decreases cholesterol absorption in theintestine is ezetimibe.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least anti-PCSK9 antibody or fragment accordingto the invention, simultaneously or sequentially with an agent thatdecreases cholesterol levels. In some embodiments, the agent thatdecreases cholesterol levels is selected from the group consisting ofcertain anti-psychotic agents, certain HIV protease inhibitors, dietaryfactors such as high fructose, sucrose, cholesterol or certain fattyacids and certain nuclear receptor agonists and antagonists for RXR,RAR, LXR, FXR.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent thatincreases HDL levels. In some embodiments, the agent that increases HDLlevels is niacin, also known as nicotinic acid. In some embodiments, theniacin is a slow-release formulation.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat inhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions. In some embodiments, the agent thatinhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions is acipimox.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat decreases lipid absorption in the intestine. In some embodiments,the agent that decreases lipid absorption in the intestine is selectedfrom the group consisting of orlistat, lipstatin, and some combinationthereof or another known in the art.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat is an anti-hypertensive and/or treats angina. In some embodiments,the agent that is an anti-hypertensive and/or treats angina isamlodipine or another anti-hypertensive such as those disclosed hereinand known in the art.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with at leastone other agent, wherein the combination allows for mitigation ofundesirable side-effects of at least one other agent. In someembodiments, the antibody or antibody fragment according to theinvention and the at least one other agent are administeredconcurrently. In some embodiments, the anti-PCSK9 antibody or fragmentaccording to the invention and at least one other agent are notadministered simultaneously, with the antibody or antibody fragmentaccording to the invention being administered before or after the atleast one other agent is administered.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one anti-PCSK9 antibody or fragment according to theinvention as provided herein.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that elevates theavailability of LDLR protein. In some embodiments, the agent thatelevates the availability of LDLR protein levels comprises a statin. Insome embodiments, the statin is selected from the group consisting ofatorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin,pitavastatin, pravastatin, rosuvastatin, simvastatin, and somecombination thereof or another statin known in the art. In someembodiments, the statin is combined with niacin, an absorption inhibitor(ezetimibe), a lipid modifying agent, or a combination thereof. In someembodiments, the agent that elevates the availability of LDLR proteinlevels comprises certain cytokines like oncostatin M, estrogen, and/orcertain herbal ingredients such as berberine.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that increaseshigh density lipoprotein (HDL) and/or decreases triglyceride levels. Insome embodiments, the agent that increases high density lipoprotein(HDL) and/or decreases triglyceride levels comprises a fibrate. In someembodiments, the fibrate is selected from the group consisting ofbezafibrate, ciprofibrate, clofibrate, gemfibrozil, fenofibrate, andsome combination thereof.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with a bile acid sequesteringagent. In some embodiments, the bile acid sequestering agent is selectedfrom the group consisting of cholestyramine, colesevelam, colestipol,and some combination thereof.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that decreasescholesterol absorption in the intestine. In some embodiments, the agentthat decreases cholesterol absorption in the intestine is ezetimibe.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that decreasescholesterol levels. In some embodiments, the agent that decreasescholesterol levels is selected from the group consisting of certainanti-psychotic agents, certain HIV protease inhibitors, dietary factorssuch as high fructose, sucrose, cholesterol or certain fatty acids andcertain nuclear receptor agonists and antagonists for RXR, RAR, LXR,FXR.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that increasesHDL levels. In some embodiments, the agent that increases HDL levels isniacin, also known as nicotinic acid. In some embodiments, the niacin isa slow-release formulation.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that inhibitshepatic triglyceride production and/or very low density lipoprotein(VLDL) secretions. In some embodiments, the agent that inhibits hepatictriglyceride production and/or very low density lipoprotein (VLDL)secretions is acipimox.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that decreaseslipid absorption in the intestine. In some embodiments, the agent thatdecreases lipid absorption in the intestine is selected from the groupconsisting of orlistat, lipstatin, and some combination thereof.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that is ananti-hypertensive and/or treats angina. In some embodiments, the agentthat is an anti-hypertensive and/or treats angina is amlodipine.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with at least one other agent,wherein the combination allows for mitigation of undesirableside-effects of at least one other agent. In some embodiments, theantibody or antibody fragment according to the invention and the atleast one other agent are administered concurrently. In someembodiments, the antibody or antibody fragment according to theinvention and at least one other agent are not administeredsimultaneously, with the antibody or antibody fragment according to theinvention being administered before or after the at least one otheragent is administered.

In some aspects, the invention comprises a neutralizing antibody orantibody fragment that binds to PCSK9 and reduces the low densitylipoprotein receptor (LDLR) lowering effect of PCSK9 on LDLR. In someembodiments, the antibody or antibody fragment specifically binds toPCSK9. In some embodiments, the antibody or fragment binds to thecatalytic domain of PCSK9.

In another embodiment of the invention humanized versions of theinventive antibodies may be derived from rabbit antibodies, using thehumanization methods disclosed herein or other known humanizationmethods.

In particular, the invention contemplates any anti-PCSK9 antibody thatcontains at least one, two, three, four, five or all six of the CDRs ofthe anti-PCSK9 antibodies exemplified herein, as well as anti-PCSK9antibodies and antibody fragments containing VH and/or VL regions whichare at least 80, 90, 95, 96, 97, 98 or 99% identical to the VH and/or VLregions of any of the anti-PCSK9 antibodies or antibody fragmentsexemplified herein. In addition the invention contemplatespolynucleotides encoding any anti-PCSK9 antibody that contains at leastone, two, three, four, five or all six of the CDRs of the anti-PCSK9antibodies exemplified herein, as well as anti-PCSK9 antibodies andantibody fragments containing VH and/or VL regions which are at least80, 90, 95, 96, 97, 98 or 99% identical to the VH and/or VL regions ofany of the anti-PCSK9 antibodies or antibody fragments exemplifiedherein, and host cells containing, i.e., antibodies or fragments capableof binding to PCSK9 and/or PCSK9/LDLR complexes.

The invention also contemplates conjugates of anti-PCSK9 antibodies andbinding fragments thereof which may conjugated to one or more effectormoiety, e.g., a detectable moiety. The invention also contemplatesmethods of making anti-PCSK9 antibodies and fragments including, but arenot limited to, Fab, Fab′, F(ab′)2, Fv, scFv fragments, SMIPs (smallmolecule immunopharmaceuticals), camelbodies, nanobodies, monovalentantibodies such as MetMabs and IgNAR.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising at least one isolated antibody or antibodyfragment according to the invention. In some embodiments, thepharmaceutical or diagnostic composition further comprises apharmaceutically acceptable diluent, carrier, solubilizer, emulsifier,preservative, or a mixture thereof. In some embodiments, thepharmaceutical or diagnostic composition further comprises at least onetherapeutic agent for inflammation. In certain embodiments, thetherapeutic agent for inflammation comprises a cyclooxygenase type 1inhibitor, a cyclooxygenase type 2 inhibitor, a small molecule modulatorof p38-MAPK, a small molecule modulator of intracellular moleculesinvolved in inflammation pathways, or mixtures thereof. In otherembodiments, the pharmaceutical or diagnostic composition islyophilized.

In some aspects, the invention comprises a method of making any of theantibody or antibody fragments containing any of the VH, VL or CDR aminoacid sequences disclosed herein in a recombinant host cell, e.g., ayeast, fungi, mammalian cell, insect cell, bacterium, avian cell or egg,or a plant cell, and preferably a yeast, filamentous fungus or mammaliancell.

This includes by way of example bacterial, fungal, yeast, mammalian,insect, plant and avian cells. Preferred host cells are yeast, fungi,especially filamentous fungi and mammalian cells. Yeast and filamentousfungi include, but are not limited to Pichia pastoris, Pichiafinlandica, Pichia trehalophila, Pichia koclamae, Pichiamembranaefaciens, Pichia minuta (Ogataea minuta, Pichia lindneri),Pichia opuntiae, Pichia thermotolerans, Pichia salictaria, Pichiaguercuum, Pichia pijperi, Pichia stiptis, Pichia methanolica, Pichiasp., Saccharomyces cerevisiae, Saccharomyces sp., Hansenula polymorpha,Kluyveromyces sp., Kluyveromyces lactis, Candida albicans, Aspergillusnidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei,Chrysosporium lucknowense, Fusarium sp., Fusarium gramineum, Fusariumvenenatum, Physcomitrella patens and Neurospora crassa. Pichia sp., anySaccharomyces sp., Hansenula polymorpha, any Kluyveromyces sp., Candidaalbicans, any Aspergillus sp., Trichoderma reesei, Chrysosporiumlucknowense, any Fusarium sp. and Neurospora crassa.

Examples of invertebrate cells include insect cells such as DrosophilaS2 and Spodoptera Sf9, as well as plant cells. Examples of usefulmammalian host cell lines include Chinese hamster ovary (CHO) and COScells. More specific examples include monkey kidney CV1 line transformedby SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293cells subcloned for growth in suspension culture, Graham et al., J. GenVirol., 36:59 (1977)); Chinese hamster ovary cells/−DHFR(CHO, Urlaub andChasin, Proc. Natl. Acad. Sci. USA, 77:4216 (1980)); mouse sertoli cells(TM4, Mather, Biol. Reprod., 23:243-251 (1980)); human lung cells (W138,ATCC CCL 75); human liver cells (Hep G2, HB 8065); and mouse mammarytumor (MMT 060562, ATCC CCL51). The selection of the appropriate hostcell is deemed to be within the skill in the art. Preferred mammaliancells for antibody expression include CHO cells and COS cells. In anexemplary embodiment the recombinant host cells are polyploidal yeastcells of the genus Pichia.

In an exemplary embodiment the host cell will comprise a Pichia pastoriscell, preferably diploidal. In preferred embodiment the host cell willsecrete the anti-PCSK9 antibody or fragment into the culture mediumcontaining host cells, e.g., Pichia pastoris, at concentrations whichare at least 10-25 mg/liter of said antibody.

Methods for producing recombinant host cells, e.g., yeast, fungi,mammalian cells, insect cells, plant cells, avian cells, bacteria, etal., that contain heterologous antibody coding sequences and whichexpress and secrete functional antibodies are well known. In anexemplary embodiment the host cells that express the antibodies orfragments according to the invention are diploidal Pichia pastoris.However, other yeast or fungi, including haploid forms, may be used.

If using diploid Pichia, these methods in general comprise producing arecombinant diploidal yeast or fungal cell that expresses and secretesthe antibody by: (i) introducing at least one expression vectorcontaining one or more heterologous polynucleotides encoding thevariable regions and optionally the constant regions of said antibody orantibody fragment, which sequences are operably linked to a promoter anda signal sequence into a host cell, e.g., a haploid yeast or fungalcell; (ii) producing by mating or spheroplast fusion a polyploidal yeastor fungi from said first and/or second haploid yeast or fungal cells;(iii) selecting polyploidal yeast or fungal cells that stably expresssaid antibody; and (iv) producing stable polyploidal yeast or fungalcultures from said polyploidal yeast or fungal cells that stably expresssaid antibody into the culture medium.

However, the yeast used may include any of the following genera:Arxiozyma; Ascobotryozyma; Citeromyces; Debaryomyces; Dekkera;Eremothecium; Issatchenkia; Kazachstania; Kluyveromyces; Kodamaea;Lodderomyces; Pachysolen; Pichia; Saccharomyces; Saturnispora;Tetrapisispora; Torulaspora; Williopsis; or Zygosaccharomyces. Incertain embodiments, the yeast genera is Pichia. In certain embodiments,the species of Pichia is selected from Pichia pastoris, Pichiamethanolica or Hansenula polymorpha (Pichia angusta). Suitablefilamentous fungi which may be used to express the subject antibodies orfragments are identified supra.

More specifically, the invention contemplates methods of making anyanti-PCSK9 antibody or fragment containing any of the VH, VL or CDRamino acid sequences disclosed herein in a recombinant cell, wherein therecombinant cell comprises a heterologous polynucleotide encoding ananti-PCSK9 antibody containing a VH amino acid sequence at least 80, 85,90, 95, 96, 97, 98 or 99% identical to any of the VH amino acidsequences in SEQ ID NO: 12, SEQ ID NO: 52, SEQ ID NO: 92, SEQ ID NO:132, SEQ ID NO: 172, SEQ ID NO: 212, SEQ ID NO: 252, SEQ ID NO: 292, SEQID NO: 332, SEQ ID NO: 372, SEQ ID NO: 412, SEQ ID NO: 452, SEQ ID NO:492, SEQ ID NO: 532, SEQ ID NO: 572, SEQ ID NO: 612, SEQ ID NO: 652, SEQID NO: 692; SEQ ID NO: 732, SEQ ID NO: 772, SEQ ID NO: 812, SEQ ID NO:852, SEQ ID NO: 892, or SEQ ID NO: 932 or encoding a variant thereofwherein at least one framework residue (FR residue) or CDR residue hasbeen altered, e.g., substituted with an amino acid present at thecorresponding position in a rabbit anti-PCSK9 antibody VH polypeptide ora conservative amino acid substitution.

In other embodiments, the heterologous polynucleotide further comprisesa polynucleotide sequence encoding an anti-PCSK9 VL amino acid sequenceat least 80, 85, 90, 95, 96, 97, 98 or 99% identical to any of the VLamino acid sequences in SEQ ID NO: 32, SEQ ID NO: 72, SEQ ID NO: 112,SEQ ID NO: 152, SEQ ID NO: 192, SEQ ID NO: 232, SEQ ID NO: 272, SEQ IDNO: 312, SEQ ID NO: 352, SEQ ID NO: 392, SEQ ID NO: 432, SEQ ID NO: 472,SEQ ID NO: 512, SEQ ID NO: 542, SEQ ID NO: 582, SEQ ID NO: 622, SEQ IDNO: 662, SEQ ID NO: 702, SEQ ID NO: 742, SEQ ID NO: 782, SEQ ID NO: 822,SEQ ID NO: 862, SEQ ID NO: 902, or SEQ ID NO: 942 or encoding a variantthereof wherein at least one framework residue (FR residue) or CDRresidue has been altered, e.g., substituted with an amino acid presentat the corresponding position in a rabbit anti-PCSK9 antibody VLpolypeptide or a conservative amino acid substitution. In otherembodiments, the heterologous polynucleotide comprises a sequenceencoding VH and VL polypeptides at least 80, 85, 90, 95, 96, 97, 98 or99% identical to any of the VH and VL amino acid sequences in containedin SEQ ID NO: 12 and SEQ ID NO: 32; SEQ ID NO: 52 and SEQ ID NO: 72; SEQID NO: 92 and SEQ ID NO: 112; SEQ ID NO: 132 and SEQ ID NO: 152; SEQ IDNO: 172 and SEQ ID NO: 192; SEQ ID NO: 212 and SEQ ID NO: 232; SEQ IDNO: 252 and SEQ ID NO: 272; SEQ ID NO: 292 and SEQ ID NO: 312; SEQ IDNO: 332 and SEQ ID NO: 352; SEQ ID NO: 372 and SEQ ID NO: 392; SEQ IDNO: 412 and SEQ ID NO: 432; SEQ ID NO: 452 and SEQ ID NO: 472; SEQ IDNO: 492 and SEQ ID NO: 512; SEQ ID NO: 532 and SEQ ID NO: 552; SEQ IDNO: 572 and SEQ ID NO: 592; SEQ ID NO: 612 and SEQ ID NO: 632; SEQ IDNO: 652 and SEQ ID NO: 672; SEQ ID NO: 692 and SEQ ID NO: 712; SEQ IDNO: 732 and SEQ ID NO: 752; SEQ ID NO: 772 and SEQ ID NO: 792; SEQ IDNO: 812 and SEQ ID NO: 832; SEQ ID NO: 852 and SEQ ID NO: 872; SEQ IDNO: 892 and SEQ ID NO: 912; SEQ ID NO: 932 and SEQ ID NO: 952; or anycombination of VH and VL sequences which are at least 80, 85, 90, 95,96, 97, 98 or 99% identical to any of the VH and VL amino acid sequencesexemplified herein.

In other embodiments, the heterologous polynucleotide encodes ananti-PCSK9 antibody or fragment wherein the antibody or fragmentcontains at least one CDR, at two or all three of the CDRs contained ina VL polypeptide and/or at least one CDR, two or all three of the CDRscontained in a VH polypeptide, wherein said VH and VL polypeptidesequences are selected from those in SEQ ID NO: 12, SEQ ID NO: 32; SEQID NO: 52, SEQ ID NO: 72; SEQ ID NO: 92, SEQ ID NO: 112; SEQ ID NO: 132,SEQ ID NO: 152; SEQ ID NO: 172, SEQ ID NO: 192; SEQ ID NO: 212, SEQ IDNO: 232, SEQ ID NO: 252, SEQ ID NO: 272; SEQ ID NO: 292, SEQ ID NO: 312;SEQ ID NO: 332, SEQ ID NO: 352; SEQ ID NO: 372, SEQ ID NO: 392; SEQ IDNO: 412, SEQ ID NO: 432; SEQ ID NO: 452, SEQ ID NO: 472; SEQ ID NO: 492,SEQ ID NO: 512; SEQ ID NO: 532; SEQ ID NO: 552; SEQ ID NO: 572; SEQ IDNO: 592; SEQ ID NO: 612; SEQ ID NO: 632; SEQ ID NO: 652; SEQ ID NO: 672;SEQ ID NO: 692; SEQ ID NO: 712; SEQ ID NO: 732; SEQ ID NO: 752; SEQ IDNO: 772; SEQ ID NO: 792; SEQ ID NO: 812; SEQ ID NO: 832; SEQ ID NO: 852;SEQ ID NO: 872; SEQ ID NO: 892; SEQ ID NO: 912; SEQ ID NO: 932; SEQ IDNO: 952; or mixtures thereof.

In some aspects, the invention comprises an isolated polynucleotidecomprising a polynucleotide encoding an anti-PCSK9 VH antibody aminoacid sequence at least 80, 85, 90, 95, 96, 97, 98, or 99% identical oridentical to the VH polypeptide sequences in SEQ ID NO: 12, SEQ ID NO:52, SEQ ID NO: 92, SEQ ID NO: 132, SEQ ID NO: 172, SEQ ID NO: 212, SEQID NO: 252, SEQ ID NO: 292, SEQ ID NO: 332, SEQ ID NO: 372, SEQ ID NO:412, SEQ ID NO: 452, SEQ ID NO: 492, SEQ ID NO: 532, SEQ ID NO: 572, SEQID NO: 612, SEQ ID NO: 652, SEQ ID NO: 692, SEQ ID NO: 732, SEQ ID NO:772, SEQ ID NO: 812, SEQ ID NO: 852, SEQ ID NO: 892, or SEQ ID NO: 932,or encoding a variant thereof wherein at least one framework residue (FRresidue) or CDR residue has been altered, e.g., substituted with anamino acid present at the corresponding position in a rabbit anti-PCSK9antibody VH polypeptide or a conservative amino acid substitution. Inother embodiments, the invention comprises a vector comprising thepolynucleotide sequence above. In other embodiments, the inventioncomprises a host cell comprising the vector mentioned above. In certainembodiments, the host cell mentioned above is a yeast cell belonging tothe genus Pichia.

In some aspects, the invention comprises an isolated polynucleotidecomprising the polynucleotide sequence encoding an anti-PCSK9 VLantibody amino acid sequence at least 80, 85, 90, 95, 96, 97, 98 or 99%identical or identical to any one of those in SEQ ID NO: 32, SEQ ID NO:72, SEQ ID NO: 112, SEQ ID NO: 152, SEQ ID NO: 192, SEQ ID NO: 232, SEQID NO: 272, SEQ ID NO: 312, SEQ ID NO: 352, SEQ ID NO: 392, SEQ ID NO:432, SEQ ID NO: 472, SEQ ID NO: 512, SEQ ID NO: 552, or SEQ ID NO: 592,SEQ ID NO: 632, SEQ ID NO: 672, SEQ ID NO: 712, SEQ ID NO: 752, SEQ IDNO: 792, SEQ ID NO: 832, SEQ ID NO: 872, SEQ ID NO: 912, or SEQ ID NO:952, or encoding a variant thereof wherein at least one frameworkresidue (FR residue) or CDR residue has been altered, e.g., substitutedwith an amino acid present at the corresponding position in a rabbitanti-PCSK9 antibody VL polypeptide or a conservative amino acidsubstitution. In other embodiments, the invention comprises a vectorcomprising any combination of the polynucleotide sequence above. Incertain embodiments, the invention comprises a host cell comprising oneor more vectors containing any of the polynucleotides mentioned above.In certain embodiments, the invention embraces a host cell containingany of the afore-mentioned polynucleotides or a vector containing, e.g.,a yeast, fungal, insect, plant, avian, bacterial or mammalian cell, andin an exemplary embodiment a yeast cell belonging to the genus Pichia.

In some aspects, the invention comprises an isolated nucleic acidcomprising a polynucleotide that hybridizes under stringent conditionsto SEQ ID NO: 12, SEQ ID NO: 32; SEQ ID NO: 52, SEQ ID NO: 72; SEQ IDNO: 92, SEQ ID NO: 112; SEQ ID NO: 132, SEQ ID NO: 152; SEQ ID NO: 172,SEQ ID NO: 192; SEQ ID NO: 212, SEQ ID NO: 232, SEQ ID NO: 252, SEQ IDNO: 272; SEQ ID NO: 292, SEQ ID NO: 312; SEQ ID NO: 332, SEQ ID NO: 352;SEQ ID NO: 372, SEQ ID NO: 392; SEQ ID NO: 412; SEQ ID NO: 432; SEQ IDNO: 452; SEQ ID NO: 472; SEQ ID NO: 492; SEQ ID NO: 512; SEQ ID NO: 532;SEQ ID NO: 552; SEQ ID NO: 572; SEQ ID NO: 592; SEQ ID NO: 612; SEQ IDNO: 632; SEQ ID NO: 652; SEQ ID NO: 672; SEQ ID NO: 692; SEQ ID NO: 712;SEQ ID NO: 732; SEQ ID NO: 752; SEQ ID NO: 772; SEQ ID NO: 792; SEQ IDNO: 812; SEQ ID NO: 832; SEQ ID NO: 852; SEQ ID NO: 872; SEQ ID NO: 892;SEQ ID NO: 912; SEQ ID NO: 932; SEQ ID NO: 952; a codon degeneratethereof; or mixtures thereof, which encodes the VH or VL region of anantibody that binds PCSK9.

The invention also pertains to the use of anti-PCSK9 antibodies andbinding fragments thereof containing or encoded by any of theafore-mentioned amino acid or polynucleotides sequences and variantsthereof for the diagnosis, assessment and treatment of any of thediseases and disorders disclosed herein, i.e., especially disordersassociated with PCSK9 or aberrant expression thereof.

The invention also contemplates the use of fragments of anti-PCSK9antibodies and binding fragments thereof containing or encoded by any ofthe afore-mentioned amino acid or polynucleotides sequences and variantsthereof for the diagnosis, assessment and treatment of diseases anddisorders associated with PCSK9 or aberrant expression thereof. Otherspecific embodiments of the invention relate to the production ofanti-PCSK9 antibodies or fragments thereof in mammalian cells such asCHO, NSO or HEK 293 cells, or yeast cells (for example diploid yeastsuch as diploid Pichia) and other yeast strains.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

FIGS. 1A-1G provide the polypeptide sequences of the full-length heavychain for antibodies Ab1-Ab24 aligned by their framework regions (FR),complementarity determining regions (CDRs), and constant regions.

FIGS. 2A-2D provide the polypeptide sequences of the full-length lightchain for antibodies Ab1-Ab24 aligned by their framework regions (FR),complementarity determining regions (CDRs), and constant regions.

FIG. 3A-3S provide the polynucleotide sequences encoding the full-lengthheavy chain for antibodies Ab1-Ab24 aligned by their framework regions(FR), complementarity determining regions (CDRs), and constant regions.

FIG. 4A-4J provide the polynucleotide sequences encoding the full-lengthlight chain for antibodies Ab1-Ab24 aligned by their framework regions(FR), complementarity determining regions (CDRs), and constant regions.

FIG. 5 provides the polypeptide sequence coordinates for the variableregion and complementarity determining regions (CDRs) of the heavy chainfor antibodies Ab1-Ab24.

FIG. 6 provides the polypeptide sequence coordinates for the constantregion and framework regions (FR) of the heavy chain for antibodiesAb1-Ab24.

FIG. 7 provides the polypeptide sequence coordinates for the variableregion and complementarity determining regions (CDRs) of the light chainfor antibodies Ab1-Ab24.

FIG. 8 provides the polypeptide sequence coordinates for the constantregion and framework regions (FR) of the light chain for antibodiesAb1-Ab24.

FIG. 9 provides the polynucleotide sequence coordinates for the variableregion and complementarity determining regions (CDRs) of the heavy chainfor antibodies Ab1-Ab24.

FIG. 10 provides the polynucleotide sequence coordinates for theconstant region and framework regions (FR) of the heavy chain forantibodies Ab1-Ab24.

FIG. 11 provides the polynucleotide sequence coordinates for thevariable region and complementarity determining regions (CDRs) of thelight chain for antibodies Ab1-Ab24.

FIG. 12 provides the polynucleotide sequence coordinates for theconstant region and framework regions (FR) of the light chain forantibodies Ab1-Ab24.

FIG. 13 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodiesAb1, Ab2, Ab3, and Ab4.

FIG. 14 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodiesAb5, Ab6, Ab8, and Ab9.

FIG. 15 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodiesAb7, Ab10, and Ab13.

FIG. 16 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodiesAb11 and Ab12.

FIG. 17 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodyAb14.

FIG. 18 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodyAb15.

FIG. 19 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodiesAb16 and Ab17.

FIG. 20 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodyAb18.

FIG. 21 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodyAb19.

FIG. 22 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodyAb20.

FIG. 23 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodyAb21.

FIG. 24 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodyAb22.

FIG. 25 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodyAb23.

FIG. 26 provides the binding data for anti-PCSK9 antibodies to humanPCSK9, obtained following the protocol in Example 1 infra for antibodyAb24.

FIG. 27 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab1.

FIG. 28 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab2.

FIG. 29 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab3.

FIG. 30 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab4.

FIG. 31 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab5.

FIG. 32 provides uptake inhibition data obtained following the protocolin Example 1 infra for antibodies Ab6 and Ab7.

FIG. 33 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab8.

FIG. 34 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab9.

FIG. 35 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibodies Ab10, Ab11, and Ab12.

FIG. 36 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab13.

FIG. 37 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab14.

FIG. 38 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab15.

FIG. 39 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibodies Ab16 and Ab17.

FIG. 40 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab18.

FIG. 41 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab19.

FIG. 42 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab20.

FIG. 43 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab21.

FIG. 44 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab22.

FIG. 45 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab23.

FIG. 46 provides LDL-C uptake inhibition data obtained following theprotocol in Example 1 infra for antibody Ab24.

FIG. 47 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibodies Ab1, Ab2, Ab3, and Ab4.

FIG. 48 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibodies Ab5, Ab6, Ab7, and Ab8.

FIG. 49 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibodies Ab9, Ab10, Ab11, and Ab12.

FIG. 50 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab13.

FIG. 51 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab14.

FIG. 52 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab15.

FIG. 53 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibodies Ab16 and Ab17.

FIG. 54 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab18.

FIG. 55 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab19.

FIG. 56 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab20.

FIG. 57 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab21.

FIG. 58 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab22.

FIG. 59 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab23.

FIG. 60 provides the binding data for anti-PCSK9 antibodies tocynomolgus monkey PCSK9, obtained following the protocol in Example 1infra for antibody Ab24.

FIG. 61 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab1.

FIG. 62 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab2.

FIG. 63 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab3.

FIG. 64 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab4.

FIG. 65 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab5.

FIG. 66 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab6.

FIG. 67 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab7.

FIG. 68 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab8.

FIG. 69 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab9.

FIG. 70 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab10.

FIG. 71 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab11.

FIG. 72 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab12.

FIG. 73 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab13.

FIG. 74 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab14.

FIG. 75 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab15.

FIG. 76 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab16.

FIG. 77 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab17.

FIG. 78 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab18.

FIG. 79 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab19.

FIG. 80 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab20.

FIG. 81 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab21.

FIG. 82 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab22.

FIG. 83 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab23.

FIG. 84 provides the inhibition data for anti-PCSK9 antibodies to blockthe interaction of PCSK9 with LDLR, obtained following the protocol inExample 6 infra for antibody Ab24.

FIG. 85 provides data showing the serum LDL-C cholesterol level incynomolgus monkeys injected with Ab18.

FIG. 86 provides data showing the serum total cholesterol level incynomolgus monkeys injected with Ab18 as a percentage change frompre-dose levels.

FIG. 87 provides data showing the serum HDL cholesterol level incynomolgus monkeys injected with Ab18 as a percentage change frompre-dose levels.

FIG. 88 provides data showing the serum LDL-C cholesterol level incynomolgus monkeys injected with Ab11 as a percentage change frompre-dose levels.

FIG. 89 provides data showing the serum total cholesterol level incynomolgus monkeys injected with Ab11 as a percentage change frompre-dose levels.

FIG. 90 provides data showing the serum HDL cholesterol level incynomolgus monkeys injected with Ab11 as a percentage change frompre-dose levels.

FIG. 91 provides data showing the serum LDL-C cholesterol level incynomolgus monkeys injected with Ab19 as a percentage change frompre-dose levels.

FIG. 92 provides data showing the serum LDL-C cholesterol level incynomolgus monkeys injected with Ab20 as a percentage change frompre-dose levels.

FIG. 93 provides data showing the serum LDL-C cholesterol level incynomolgus monkeys injected with Ab22 as a percentage change frompre-dose levels.

FIG. 94 provides data showing the serum LDL-C cholesterol level incynomolgus monkeys injected with Ab24 as a percentage change frompre-dose levels.

PREFERRED EMBODIMENTS OF THE INVENTION

Different preferred embodiments of the invention are listed below andare further described in more detail in the Detailed Written Descriptionand in the Examples section of this application.

A preferred embodiment of the invention relates to an anti-human PCSK9antibody or antibody fragment that competes with and/or specificallybinds to the same or overlapping epitope(s) on human PCSK9 as ananti-human PCSK9 antibody selected from the group consisting of Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14,Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23 and Ab24.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein said antibody or antibodyfragment pacifically binds to the same or overlapping epitope(s) onhuman PCSK9 as an anti-human PCSK9 antibody selected from the groupconsisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11,Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23and Ab24.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein said epitope(s) areidentified using a binding assay that detects the binding of saidanti-human PCSK9 antibody to one or more peptides in a library of10-15-mer peptides which are overlapping peptide fragments of humanPCSK9 that correspond to all or substantially all of the length of thehuman PCSK9 polypeptide.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein said binding assay is aWestern immunoblot assay which detects specific binding of the antibodyor antibody fragment to one or more of said 10-15 mer peptides in saidlibrary by the use of a chemiluminescent label which emits a detectablechemiluminescent signal when specific binding of said antibody orantibody fragment to a peptide in said library occurs.

Another preferred embodiment of the invention relates to an anti-humanantibody PCSK9 antibody or antibody fragment wherein said antibody orantibody fragment contains at least 2 complementarity determiningregions (CDRs) of an anti-PCSK9 antibody selected from the groupconsisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11,Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23and Ab24.

Another preferred embodiment of the invention relates to an anti-humanantibody PCSK9 antibody or antibody fragment, wherein said antibody orantibody fragment contains at least 3 complementarity determiningregions (CDRs) of an anti-PCSK9 antibody selected from the groupconsisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11,Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23and Ab24.

Another preferred embodiment of the invention relates to an anti-humanantibody PCSK9 antibody or antibody fragment, wherein said antibody orantibody fragment contains at least 4 complementarity determiningregions (CDRs) of an anti-PCSK9 antibody selected from the groupconsisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11,Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23and Ab24.

Another preferred embodiment of the invention relates to an anti-humanantibody PCSK9 antibody or antibody fragment, wherein said antibody orantibody fragment contains at least 5 complementarity determiningregions (CDRs) of an anti-PCSK9 antibody selected from the groupconsisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11,Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23and Ab24.

Another preferred embodiment of the invention relates to an anti-humanantibody PCSK9 antibody or antibody fragment, wherein said antibody orantibody fragment contains all 6 complementarity determining regions(CDRs) of an anti-PCSK9 antibody selected from the group consisting ofAb1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13,Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23 and Ab24.

Another preferred embodiment of the invention relates to an anti-humanantibody PCSK9 antibody or antibody fragment wherein said antibody orantibody fragment contains (a) a variable heavy chain comprising a CDR1sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO:44, SEQ ID NO: 84, SEQ ID NO: 124, SEQ ID NO: 164, SEQ ID NO: 204, SEQID NO: 244, SEQ ID NO: 284, SEQ ID NO: 324, SEQ ID NO: 364, SEQ ID NO:404, SEQ ID NO: 444, SEQ ID NO: 484, SEQ ID NO: 524, SEQ ID NO: 564, SEQID NO: 604, SEQ ID NO: 644, SEQ ID NO: 684, SEQ ID NO: 724, SEQ ID NO:764, SEQ ID NO: 804, SEQ ID NO: 844, SEQ ID NO: 884, and SEQ ID NO: 924;a CDR2 sequence selected from the group consisting of SEQ ID NO: 6, SEQID NO: 46, SEQ ID NO: 86, SEQ ID NO: 126, SEQ ID NO: 166, SEQ ID NO:206, SEQ ID NO: 246, SEQ ID NO: 286, SEQ ID NO: 326, SEQ ID NO: 366, SEQID NO: 406, SEQ ID NO: 446, SEQ ID NO: 486, SEQ ID NO: 526, SEQ ID NO:566, SEQ ID NO: 606, SEQ ID NO: 646, SEQ ID NO: 686, SEQ ID NO: 726, SEQID NO: 766, SEQ ID NO: 806, SEQ ID NO: 846, SEQ ID NO: 886, and SEQ IDNO: 926; and a CDR3 sequence selected from the group consisting of SEQID NO: 8, SEQ ID NO: 48, SEQ ID NO: 88, SEQ ID NO: 128, SEQ ID NO: 168,SEQ ID NO: 208, SEQ ID NO: 248, SEQ ID NO: 288, SEQ ID NO: 328, SEQ IDNO: 368, SEQ ID NO: 408, SEQ ID NO: 448, SEQ ID NO: 488, SEQ ID NO: 528,SEQ ID NO: 568, SEQ ID NO: 608, SEQ ID NO: 648, SEQ ID NO: 688, SEQ IDNO: 728, SEQ ID NO: 768, SEQ ID NO: 808, SEQ ID NO: 848, SEQ ID NO: 888,and SEQ ID NO: 928; and/or

-   -   (b) a variable light chain comprising a CDR1 sequence selected        from the group consisting of SEQ ID NO: 24, SEQ ID NO: 64, SEQ        ID NO: 104, SEQ ID NO: 144, SEQ ID NO: 184, SEQ ID NO: 224, SEQ        ID NO: 264, SEQ ID NO: 304, SEQ ID NO: 344, SEQ ID NO: 384, SEQ        ID NO: 424, SEQ ID NO: 464, SEQ ID NO: 504, SEQ ID NO: 544, SEQ        ID NO: 584, SEQ ID NO: 624, SEQ ID NO: 664, SEQ ID NO: 704, SEQ        ID NO: 744, SEQ ID NO: 784, SEQ ID NO: 824, SEQ ID NO: 864, SEQ        ID NO: 904, and SEQ ID NO: 944; a CDR2 sequence selected from        the group consisting of SEQ ID NO: 26, SEQ ID NO: 66, SEQ ID NO:        106, SEQ ID NO: 146, SEQ ID NO: 186, SEQ ID NO: 226, SEQ ID NO:        266, SEQ ID NO: 306, SEQ ID NO: 346, SEQ ID NO: 386, SEQ ID NO:        426, SEQ ID NO: 466, SEQ ID NO: 506, SEQ ID NO: 546, SEQ ID NO:        586, SEQ ID NO: 626, SEQ ID NO: 666, SEQ ID NO: 706, SEQ ID NO:        746, SEQ ID NO: 786, SEQ ID NO: 826, SEQ ID NO: 866, SEQ ID NO:        906, and SEQ ID NO: 946; and a CDR3 sequence selected from the        group consisting of SEQ ID NO: 28, SEQ ID NO: 68, SEQ ID NO:        108, SEQ ID NO: 148, SEQ ID NO: 188, SEQ ID NO: 228, SEQ ID NO:        268, SEQ ID NO: 308, SEQ ID NO: 348, SEQ ID NO: 388, SEQ ID NO:        428, SEQ ID NO: 468, SEQ ID NO: 508, SEQ ID NO: 548, SEQ ID NO:        588, SEQ ID NO: 628, SEQ ID NO: 668, SEQ ID NO: 708, SEQ ID NO:        748, SEQ ID NO: 788, SEQ ID NO: 828, SEQ ID NO: 868, SEQ ID NO:        908, and SEQ ID NO: 948, with the further proviso that one or        two residues of any of the afore-identified CDR polypeptides may        be substituted with another amino acid, preferably a        conservative amino acid substitution.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 4, the CDR2 sequence of SEQ IDNO: 6, and the CDR3 sequence of SEQ ID NO: 8; and/or the variable lightchain comprises the CDR1 sequence of SEQ ID NO: 24, the CDR2 sequence ofSEQ ID NO: 26, and the CDR3 sequence of SEQ ID NO: 28.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 44, the CDR2 sequence of SEQID NO: 46, and the CDR3 sequence of SEQ ID NO: 48; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 64, the CDR2sequence of SEQ ID NO: 66, and the CDR3 sequence of SEQ ID NO: 68.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 84, the CDR2 sequence of SEQID NO: 86, and the CDR3 sequence of SEQ ID NO: 88; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 104, the CDR2sequence of SEQ ID NO: 106, and the CDR3 sequence of SEQ ID NO: 108.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 124, the CDR2 sequence of SEQID NO: 126, and the CDR3 sequence of SEQ ID NO: 128; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 144, the CDR2sequence of SEQ ID NO: 146, and the CDR3 sequence of SEQ ID NO: 148.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 164, the CDR2 sequence of SEQID NO: 166, and the CDR3 sequence of SEQ ID NO: 168; and the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 184, the CDR2sequence of SEQ ID NO: 186, and the CDR3 sequence of SEQ ID NO: 188.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises the CDR1 sequence of SEQ ID NO: 204,the CDR2 sequence of SEQ ID NO: 206, and the CDR3 sequence of SEQ ID NO:208; and the variable light chain comprises the CDR1 sequence of SEQ IDNO: 224, the CDR2 sequence of SEQ ID NO: 226, and the CDR3 sequence ofSEQ ID NO: 228.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 244, the CDR2 sequence of SEQID NO: 246, and the CDR3 sequence of SEQ ID NO: 248; and the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 264, the CDR2sequence of SEQ ID NO: 266, and the CDR3 sequence of SEQ ID NO: 268.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 284, the CDR2 sequence of SEQID NO: 286, and the CDR3 sequence of SEQ ID NO: 288; and the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 304, the CDR2sequence of SEQ ID NO: 306, and the CDR3 sequence of SEQ ID NO: 308.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 324, the CDR2 sequence of SEQID NO: 326, and the CDR3 sequence of SEQ ID NO: 328; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 344, the CDR2sequence of SEQ ID NO: 346, and the CDR3 sequence of SEQ ID NO: 348.

Another preferred embodiments of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 364, the CDR2 sequence of SEQID NO: 366, and the CDR3 sequence of SEQ ID NO: 368; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 384, the CDR2sequence of SEQ ID NO: 386, and the CDR3 sequence of SEQ ID NO: 388.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 404, the CDR2 sequence of SEQID NO: 406, and the CDR3 sequence of SEQ ID NO: 408; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 424, the CDR2sequence of SEQ ID NO: 426, and the CDR3 sequence of SEQ ID NO: 428.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 444, the CDR2 sequence of SEQID NO: 446, and the CDR3 sequence of SEQ ID NO: 448; and the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 464, the CDR2sequence of SEQ ID NO: 466, and the CDR3 sequence of SEQ ID NO: 468.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 484, the CDR2 sequence of SEQID NO: 486, and the CDR3 sequence of SEQ ID NO: 488; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 504, the CDR2sequence of SEQ ID NO: 506, and the CDR3 sequence of SEQ ID NO: 508.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 524, the CDR2 sequence of SEQID NO: 526, and the CDR3 sequence of SEQ ID NO: 528; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 544, the CDR2sequence of SEQ ID NO: 546, and the CDR3 sequence of SEQ ID NO: 548.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 564, the CDR2 sequence of SEQID NO: 566, and the CDR3 sequence of SEQ ID NO: 568; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 584, the CDR2sequence of SEQ ID NO: 586, and the CDR3 sequence of SEQ ID NO: 588.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 604, the CDR2 sequence of SEQID NO: 606, and the CDR3 sequence of SEQ ID NO: 608; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 624, the CDR2sequence of SEQ ID NO: 626, and the CDR3 sequence of SEQ ID NO: 628.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 644, the CDR2 sequence of SEQID NO: 646, and the CDR3 sequence of SEQ ID NO: 648; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 664, the CDR2sequence of SEQ ID NO: 666, and the CDR3 sequence of SEQ ID NO: 668.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 684, the CDR2 sequence of SEQID NO: 686, and the CDR3 sequence of SEQ ID NO: 688; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 704, the CDR2sequence of SEQ ID NO: 706, and the CDR3 sequence of SEQ ID NO: 708.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 724, the CDR2 sequence of SEQID NO: 726, and the CDR3 sequence of SEQ ID NO: 728; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 744, the CDR2sequence of SEQ ID NO: 746, and the CDR3 sequence of SEQ ID NO: 748.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 764, the CDR2 sequence of SEQID NO: 766, and the CDR3 sequence of SEQ ID NO: 768; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 784, the CDR2sequence of SEQ ID NO: 786, and the CDR3 sequence of SEQ ID NO: 788.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 804, the CDR2 sequence of SEQID NO: 806, and the CDR3 sequence of SEQ ID NO: 808; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 824, the CDR2sequence of SEQ ID NO: 826, and the CDR3 sequence of SEQ ID NO: 828.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 844, the CDR2 sequence of SEQID NO: 846, and the CDR3 sequence of SEQ ID NO: 848; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 864, the CDR2sequence of SEQ ID NO: 866, and the CDR3 sequence of SEQ ID NO: 868.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 884, the CDR2 sequence of SEQID NO: 886, and the CDR3 sequence of SEQ ID NO: 888; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 904, the CDR2sequence of SEQ ID NO: 906, and the CDR3 sequence of SEQ ID NO: 908.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises the CDR1 sequence of SEQ ID NO: 924, the CDR2 sequence of SEQID NO: 926, and the CDR3 sequence of SEQ ID NO: 928; and/or the variablelight chain comprises the CDR1 sequence of SEQ ID NO: 944, the CDR2sequence of SEQ ID NO: 946, and the CDR3 sequence of SEQ ID NO: 948.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy chaincomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 2 and/or the variable light chain comprises SEQID NO: 22.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 2 and SEQ ID NO: 22, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 42 and the variable light chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 62.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 42 and SEQ ID NO: 62, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 82 and the variable lightchain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 102.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy and lightchains are each at least 90% identical to the variable heavy and lightchains in SEQ ID NO: 82 and SEQ ID NO: 102, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 122 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 142.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy and lightchains are each at least 90% identical to the variable heavy and lightchains in SEQ ID NO: 122 and SEQ ID NO: 142, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 162 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 182.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 162 and SEQ ID NO: 182, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 202 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 222.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 202 and SEQ ID NO: 222, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 242 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 262.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy and lightchains are each at least 90% identical to the variable heavy and lightchains in SEQ ID NO: 242 and SEQ ID NO: 262, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 282 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 302.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 282 and SEQ ID NO: 302, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 322 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 342.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy and lightchains are each at least 90% identical to the variable heavy and lightchains in SEQ ID NO: 322 and SEQ ID NO: 342, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 362 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 382.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 362 and SEQ ID NO: 382, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 402 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 422.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment 55, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 402 and SEQ ID NO: 422, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 442 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 462.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 442 and SEQ ID NO: 462, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 482 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 502.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 482 and SEQ ID NO: 502, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 522 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 542.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 522 and SEQ ID NO: 542, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 562 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 582.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 562 and SEQ ID NO: 582, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 602 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 622.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 602 and SEQ ID NO: 622, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 642 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 662.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 642 and SEQ ID NO: 662, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment according to the invention, whereinthe variable heavy chain comprises a sequence at least 80, 85, 90, 95,96, 97, 98, 99 or 100% identical to SEQ ID NO: 682 and the variablelight chain comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99or 100% identical to SEQ ID NO: 702.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy and lightchains are each at least 90% identical to the variable heavy and lightchains in SEQ ID NO: 682 and SEQ ID NO: 702, respectively

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment of wherein the variable heavy chaincomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 722 and the variable light chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 742.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy and lightchains are each at least 90% identical to the variable heavy and lightchains in SEQ ID NO: 722 and SEQ ID NO: 742, respectively

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment of wherein the variable heavy chaincomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 762 and the variable light chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 782.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy and lightchains are each at least 90% identical to the variable heavy and lightchains in SEQ ID NO: 762 and SEQ ID NO: 782, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy chaincomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 802 and the variable light chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 822.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 802 and SEQ ID NO: 822, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment of wherein the variable heavy chaincomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 842 and the variable light chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 862.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the variable heavy andlight chains are each at least 90% identical to the variable heavy andlight chains in SEQ ID NO: 842 and SEQ ID NO: 862, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment of wherein the variable heavy chaincomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 882 and the variable light chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 902.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy and lightchains are each at least 90% identical to the variable heavy and lightchains in SEQ ID NO: 882 and SEQ ID NO: 902, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment of, wherein the variable heavy chaincomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 922 and the variable light chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 942.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the variable heavy and lightchains are each at least 90% identical to the variable heavy and lightchains in SEQ ID NO: 922 and SEQ ID NO: 942, respectively.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 1 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 21.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 41 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 61.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 81 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 101.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 121 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 141.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 161 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 181.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 201 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 221.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 241 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 261.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 281 and the light chain comprises SEQ ID NO: 301.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprisesSEQ ID NO: 321 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 341.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 361 and the light chain comprises SEQ ID NO: 381.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprisesSEQ ID NO: 401 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 421.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 441 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 461.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 481 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 501.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 521 and the light chain comprises SEQ ID NO: 541.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the heavy chain comprisesSEQ ID NO: 561 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 581.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 601 and the light chain comprises SEQ ID NO: 621.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprisesSEQ ID NO: 641 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 661.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment of wherein the heavy chain comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 681 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 701.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 721 and the light chain comprises SEQ ID NO: 741.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprisesSEQ ID NO: 761 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 781.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment of wherein the heavy chain comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 801 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 821.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 841 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 861.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 881 and the light chain comprises SEQ ID NO: 901.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the heavy chain comprisesSEQ ID NO: 921 and the light chain comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 941.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the antibody or antibodyfragment is selected from the group consisting of chimeric, humanized,and human antibodies or antibody fragments.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment is selected from the group consisting of scFvs, camelbodies,nanobodies, IgNAR, F_(ab) fragments, F_(ab′) fragments, MetMab likeantibodies, monovalent antibody fragments, and F_((ab′)2) fragments.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment substantially or entirely lacks N-glycosylation and/orO-glycosylation.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment comprises a human constant domain.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody is an IgG1,IgG2, IgG3, or IgG4 antibody.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment comprises an Fc region that has been modified to alter at leastone of effector function, half-life, proteolysis, or glycosylation.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the Fc region contains oneor more mutations that alter or eliminate N- and/or O-glycosylation.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment is a humanized antibody or antibody fragment.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the antibody or antibodyfragment binds to PCSK9 with a dissociation constant (K_(d)) of lessthan or equal to 10⁻² M, 5×10⁻³ M, 10⁻³ M, 5×10⁻⁴ M, 10⁻⁴ M, 5×10⁻⁵ M,10⁻⁵ M, 5×10⁻⁶ M, 10⁻⁶ M, 5×10⁻⁷ M, 10⁻⁷ M, 5×10⁻⁸ M, 10⁻⁸ M, 5×10⁻⁹ M,10⁻⁹ M, 5×10⁻¹⁰ M, 10⁻¹⁰ M, 5×10⁻¹¹ M, 10⁻¹¹ M, 5×10⁻¹² M, 10⁻¹² M,5×10⁻¹³ M, or 10⁻¹³ M.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment binds to PCSK9 with a dissociation constant (K_(d)) of lessthan or equal to 10⁻¹¹ M, 5×10⁻¹² M, or 10⁻¹² M.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, which binds to PCSK9 with anoff-rate of less than or equal to 10⁻⁴ S⁻¹, 5×10⁻⁵ S⁻¹, 10⁻⁵ S⁻¹, 5×10⁻⁶S⁻¹, 10⁻⁶ S⁻¹, 5×10⁻⁷ S⁻¹, or 10⁻⁷ S⁻¹.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment is directly or indirectly attached to a detectable label ortherapeutic agent.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, which when administered to a humansubject inhibits or neutralizes at least one biological effect elicitedby PCSK9.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment which when administered to a humansubject reduces serum cholesterol.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment is capable of inhibiting the binding of PCSK9 to LDLR.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment binds to PCSK9 with a K_(D) that is less than about 100 nM.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment binds to PCSK9 with a K_(D) that is less than about 10 nM.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment binds to PCSK9 with a K_(D) that is less than about 1 nM.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment binds to PCSK9 with a K_(D) that is between about 1 and about10 nM.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment binds to PCSK9 with a K_(D) that is between about 0.1 and about1 nM.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment binds to PCSK9 with a K_(D) that is between 0.12 and 7.99 nM.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment is attached to at least one effector moiety.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein effector moiety comprises achemical linker.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein the antibody or antibodyfragment is attached to one or more detectable moieties.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment, wherein detectable moiety comprisesa fluorescent dye, enzyme, substrate, bioluminescent material,radioactive material, chemiluminescent moiety, or mixtures thereof.

Another preferred embodiment of the invention relates to an anti-humanPCSK9 antibody or antibody fragment wherein the antibody or antibodyfragment is attached to one or more functional moieties.

An anti-idiotypic antibody produced against an anti-human PCSK9 antibodyor antibody fragment according to the invention.

A composition suitable for therapeutic, prophylaxis, or a diagnostic usecomprising a therapeutically, prophylactically or diagnosticallyeffective amount of at least one antibody or antibody fragment accordingto the invention.

Another preferred embodiment of the invention relates to an composition,which is suitable for subcutaneous administration containing at leastone antibody or antibody fragment according to the invention.

Another preferred embodiment of the invention relates to an compositionwhich is suitable for intravenous administration containing at least oneantibody or antibody fragment according to the invention.

Another preferred embodiment of the invention relates to a composition,which is suitable for topical administration containing at least oneantibody or antibody fragment according to the invention.

Another preferred embodiment of the invention relates to a composition,which is lyophilized containing at least one antibody or antibodyfragment according to the invention.

Another preferred embodiment of the invention relates to a compositioncontaining at least one antibody or antibody fragment according to theinvention further comprising a pharmaceutically acceptable diluent,carrier, solubilizer, emulsifier, preservative, or mixture thereof.

Another preferred embodiment of the invention relates to a compositioncontaining at least one antibody or antibody fragment according to theinvention, further comprising another active agent.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the other active agent is selected from the groupconsisting of the following: statins, ACE inhibitors, Angiotensin IIreceptor blockers (ARBs), Antiarrhythmics, Antiplatelet Drugs, aspirin,beta blockers, amiodarone, digoxin, aspirin, anti-clotting agents,digoxin, diuretics, heart failure drugs, vasodilators, blood thinners,other anti-cholesterol drugs such as holestyramine (Questran),gemfibrozil (Lopid, Gemcor), Omacor, pantethine, anti-hypertensives,antidiabetigenic drugs, Meglitinides, Sulfonylurea, andThiazolidinediones.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the ACE inhibitor is selected from: Capoten(captopril), Vasotec (enalapril), Prinivil, Zestril (lisinopril),Lotensin (benazepril), Monopril (fosinopril), Altace (ramipril),Accupril (quinapril), Aceon (perindopril), Mavik (trandolapril), Univasc(moexipril),

Another preferred embodiment of the invention relates to a compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the ARB is selected from: Cozaar (losartan), Diovan(valsartan), Avapro (irbesartan), Atacand (candesartan), and Micardis(telmisartan).

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the Antiarrhythmic is selected from Tambocor(flecamide), Procanbid (procainamide), Cordarone (amiodarone), andBetapace (sotalol).

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the anticlotting agent is selected from Anticlottingagents which may be used in combination with the subject anti-PCSK9antibodies and antibody fragments include: Tissue plasminogen activator(TPA), Tenecteplase, Alteplase, Urokinase, Reteplase, and Streptokinase.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention wherein the Beta blocker is selected from Sectral(acebutolol), Zebeta (bisoprolol), Brevibloc (esmolol), Inderal(propranolol), Tenormin (atenolol), Normodyne, Trandate (labetalol),Coreg (carvedilol), Lopressor, and Toprol-XL (metoprolol).

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the calcium channel blocker is selected from: Norvasc(amlodipine), Plendil (felodipine), Cardizem, Cardizem CD, Cardizem SR,Dilacor XR, Diltia XT, Tiazac (diltiazem), Calan, Calan SR, Covera-HS,Isoptin, Isoptin SR, Verelan, Verelan PM (verapamil), Adalat, Adalat CC,Procardia, Procardia XL (nifedipine), Cardene, Cardene SR (nicardipine),Sular (nisoldipine), Vascor (bepridil), and Caduet which is acombination of a statin cholesterol drug and amlodipine.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the diuretic is selected from: Lasix (furosemide),Bumex (bumetanide), Demadex (torsemide), Esidrix (hydrochlorothiazide),Zaroxolyn (metolazone), and Aldactone (spironolactone).

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the heart failure drug is selected from Dobutrex(dobutamine), and Primacor (milrinone).

Another preferred embodiment of the invention relates to an compositioncontaining an antibody or fragment according to the invention, whereinthe vasodilator is selected from Dilatrate-SR, Iso-Bid, Isonate,Isorbid, Isordil, Isotrate, Sorbitrate (isosorbide dinitrate), IMDUR(isorbide mononitrate), and BiDil (hydralazine with isosorbidedinitrate.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the blood thinner is selected Warfarin (coumadin),Heparin, Lovenox, and Fragmin.

Another preferred embodiment of the invention relates to an compositionof containing an antibody according to the invention wherein thecomposition further comprises another therapeutic agent that elevatesthe availability of LDLR protein.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the composition further comprises another therapeuticagent that blocks or inhibits cholesterol synthesis.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the composition further comprises a statin.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the statin is selected atorvastatin, cerivastatin,fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin,rosuvastatin, simvastatin, or mixtures thereof.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the composition further comprises another therapeuticagent that elevates the HDL level, or an agent which decreasestriglyceride levels, or both.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the composition further comprises a fibrate.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the fibrate comprises bezafibrate, ciprofibrate,clofibrate, gemfibrozil, fenofibrate, or mixtures thereof.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the composition further comprises a bile sequesteringagent.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein bile sequestering agent comprises cholestyramine,colesevelam, colestipol, or mixtures thereof.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the composition further comprises an agent thatdecreases cholesterol absorption in the intestines.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the agent comprises ezetimibe.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the composition comprises an agent that that inhibitshepatic triglyceride production, inhibits VLDL secretions, or both.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention wherein the agent comprises acipimox.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the composition further comprises an agent thatdecreases lipid absorption in the intestines.

Another preferred embodiment of the invention relates to an compositioncontaining an antibody according to the invention, wherein the agentcomprises orlistat, lipstatin, or mixtures thereof.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the composition further comprises an agent that is ananti-hypertensive, an agent that treats angina, or both.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein said anti-hypertensive or anti-angina agent isselected from a diuretic, an adrenergic receptor antagonist, a calciumchannel blocker, a renin inhibitor, an ACE inhibitor, an angiotensin IIreceptor antagonist, aa aldosterone antagonist, a vasodilator or analpha-2 agonist.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the diuretics include bumetanide, thacrynic acid,furosemide, and torsemide; a thiazide diuretic such as epitizide,hydrochlorothiazide or chlorothiazide, or bendroflumethiazide; athiazide-like diuretic such as indapamide, chlorthalidone, ormetolazone; a potassium-sparing diuretic such as amiloride, triamterene,or spironolactone; the adrenergic receptor antagonists include betablockers such as atenolol, metoprolol, nadolol, oxprenolol, pindolol,propranolol, or timolol; the alpha blockers include doxazosin,phentolamine, indoramin, phenoxybenzamine, prazosin, terazosin, andtolazoline; mixed alpha+beta blockers such as bucindolol, carvedilol,and labetalol; the calcium channel blockers include dihydropyridinessuch as amlodipine, felodipine, isradipine, lercanidipine, nicardipine,nifedipine, nimodipine, and nitrendipine; non-dihydropyridines such asdiltiazem and verapamil; renin inhibitors such as Aliskiren; ACEinhibitors such as captopril, enalapril, fosinopril, lisinopril,perindopril, quinapril, ramipril, trandolapril, and benazepril;angiotensin II receptor antagonists such as candesartan, eprosartan,irbesartan, losartan, olmesartan, telmisartan, and valsartan;aldosterone antagonists such as eplerenone and spironolactone;vasodilators such as sodium nitroprusside; and Alpha-2 agonists such asClonidine, Guanabenz, Methyldopa, Moxonidine, Guanethidine or Reserpine;or a nitrate such as nitroglycerin (glyceryl trinitrate),pentaerythritol tetranitrate, isosorbide dinitrate or isosorbidemononitrate.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, which further comprises at least one other agent, wherein thecombination allows for mitigation of undesirable side effects of atleast one other agent contained therein.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention which further comprises at least one therapeutic agent forinflammation.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, wherein the therapeutic agent for inflammation comprises acyclooxygenase type 1 inhibitor, a cyclooxygenase type 2 inhibitor, asmall molecule modulator of p38-MAPK, a small molecule modulator ofintracellular molecules involved in inflammation pathways, or mixturesthereof.

Another preferred embodiment of the invention relates to an compositioncontaining at least one antibody or antibody fragment according to theinvention, which is lyophilized, stabilized and/or or formulated foradministration by injection.

Another preferred embodiment of the invention relates to a method forblocking, inhibiting or neutralizing one or more biological effectsassociated with PCSK9 comprising administering to a subject in needthereof an effective amount of at least one antibody or compositionaccording to the invention.

Another preferred embodiment of the invention relates to method fortreating or preventing a condition associated with elevated serumcholesterol levels in a patient, comprising administering to a patientin need thereof an effective amount of at least one antibody accordingto the invention or a composition containing optionally with anotheractive agent.

Another preferred embodiment of the invention relates to method fortreating or preventing elevated serum cholesterol levels in a patient inneed thereof or at risk of developing elevated serum cholesterol levelsbecause of an existing condition, comprising administering to thepatient an effective amount of at least one antibody according to aneffective amount of at least one antibody according to the invention ora composition containing optionally with another active agent.

Another preferred embodiment of the invention relates to a methodwherein the condition treated is selected from the group consisting ofhypercholesterolemia, coronary heart disease, hyperlipidemia,hypertriglyceridaemia, sitosterolemia, atherosclerosis, arteriosclerosismetabolic syndrome, acute coronary syndrome, vascular inflammation,xanthoma, diabetes, obesity, hypertension, and angina.

Another preferred embodiment is a method for treating or preventing adisorder involving cholesterol or lipid homeostasis or complicationsassociated therewith, comprising administering to the patient aneffective amount of at least one antibody according to an effectiveamount of at least one antibody according to the invention or acomposition containing optionally with another active agent.

Another preferred embodiment of the invention relates to a method oftreatment wherein the treated condition is selected fromhypercholesterolemia, hyperlipidemia, hypertriglyceridaemia, andsitosterolemia using an effective amount of at least one antibodyaccording to the invention or a composition containing optionally withanother active agent.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the at least one isolated anti-human PCSK9 antibody or antibodyinhibits the binding of PCSK9 to LDLR.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the at least one isolated anti-human PCSK9 antibody or antibodyfragment results in blocking or inhibition of cholesterol synthesis.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering separately orco-administering another agent, which agent that elevates theavailability of LDLR protein or which blocks or inhibits cholesterolsynthesis.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the administration is part of a therapeutic regimen that furtherincludes the administration of at least one of the following: statins,ACE inhibitors, Angiotensin II receptor blockers (ARBs),Antiarrhythmics, Antiplatelet Drugs, aspirin, beta blockers, amiodarone,digoxin, aspirin, anti-clotting agents, digoxin, diuretics, heartfailure drugs, vasodilators, blood thinners, other anti-cholesteroldrugs such as holestyramine (Questran), gemfibrozil (Lopid, Gemcor),Omacor, pantethine, anti-hypertensives, antidiabetigenic drugs,Meglitinides, Sulfonylurea, and Thiazolidinediones.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the ACE inhibitor is selected from: Capoten (captopril), Vasotec(enalapril), Prinivil, Zestril (lisinopril), Lotensin (benazepril),Monopril (fosinopril), Altace (ramipril), Accupril (quinapril), Aceon(perindopril), Mavik (trandolapril), Univasc (moexipril),

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe ARB is selected from: Cozaar (losartan), Diovan (valsartan), Avapro(irbesartan), Atacand (candesartan), and Micardis (telmisartan).

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe antiarythmic is selected from: Tambocor (flecamide), Procanbid(procainamide), Cordarone (amiodarone), and Betapace (sotalol).

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the anti-clotting agent is selected from: Tissue plasminogenactivator (TPA), Tenecteplase, Alteplase, Urokinase, Reteplase, andStreptokinase.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe Beta-blocker is selected from: Sectral (acebutolol), Zebeta(bisoprolol), Brevibloc (esmolol), Inderal (propranolol), Tenormin(atenolol), Normodyne, Trandate (labetalol), Coreg (carvedilol),Lopressor, and Toprol-XL (metoprolol).

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the calcium channel blocker is selected from: Norvasc(amlodipine), Plendil (felodipine), Cardizem, Cardizem CD, Cardizem SR,Dilacor XR, Diltia XT, Tiazac (diltiazem), Calan, Calan SR, Covera-HS,Isoptin, Isoptin SR, Verelan, Verelan PM (verapamil), Adalat, Adalat CC,Procardia, Procardia XL (nifedipine), Cardene, Cardene SR (nicardipine),Sular (nisoldipine), Vascor (bepridil), and Caduet which is acombination of a statin cholesterol drug and amlodipine.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe antidiuretic is selected from: Lasix (furosemide), Bumex(bumetanide), Demadex (torsemide), Esidrix (hydrochlorothiazide),Zaroxolyn (metolazone), and Aldactone (spironolactone).

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the heart failure drug is selected from Dobutrex (dobutamine),and Primacor (milrinone).

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the vasodilator is selected from: Dilatrate-SR, Iso-Bid,Isonate, Isorbid, Isordil, Isotrate, Sorbitrate (isosorbide dinitrate),IMDUR (isorbide mononitrate), and BiDil (hydralazine with isosorbidedinitrate.

Another preferred embodiment of the invention relates to a methodcontaining an antibody according to the invention, wherein the bloodthinner is selected from Warfarin (coumadin), Heparin, Lovenox, andFragmin.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent that elevates the availability of LDLR protein orwhich blocks or inhibits cholesterol synthesis is administeredsimultaneously with the antibody or composition containing.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein another agent that elevates the availability of LDLR protein orwhich blocks or inhibits cholesterol synthesis is administered.sequentially relative to the administration of the antibody orcomposition containing.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent that blocks or inhibits cholesterol synthesis proteincomprises a statin.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the statin is selected from atorvastatin, cerivastatin,fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin,rosuvastatin, simvastatin, or mixtures thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe method further comprises administering an agent that elevates theHDL level, an agent that decreases triglyceride levels, or both.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises a fibrate.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the fibrate agent comprises bezafibrate, ciprofibrate,clofibrate, gemfibrozil, fenofibrate, or mixtures thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering a bile sequesteringagent.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the bile sequestering agent comprises cholestyramine,colesevelam, colestipol, or mixtures thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering an agent thatdecreases cholesterol absorption in the intestines.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises ezetimibe.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent 174,wherein the method further comprises administering an agent thatinhibits hepatic triglyceride production, inhibits VLDL secretions, orboth.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises acipimox.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering an agent thatdecreases lipid absorption in the intestines.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises orlistat, lipstatin, or mixtures thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent 174,wherein the method further comprises administering an agent that is ananti-hypertensive, one that treats angina, or both.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein said anti-hypertensive or anti-angina agent is selected from adiuretic, an adrenergic receptor antagonist, a calcium channel blocker,a renin inhibitor, an ACE inhibitor, an angiotensin II receptorantagonist, aa aldosterone antagonist, a vasodilator or an alpha-2agonist.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the diuretics include bumetanide, thacrynic acid, furosemide,and torsemide; a thiazide diuretic such as epitizide,hydrochlorothiazide or chlorothiazide, or bendroflumethiazide; athiazide-like diuretic such as indapamide, chlorthalidone, ormetolazone; a potassium-sparing diuretic such as amiloride, triamterene,or spironolactone; the adrenergic receptor antagonists include betablockers such as atenolol, metoprolol, nadolol, oxprenolol, pindolol,propranolol, or timolol; the alpha blockers include doxazosin,phentolamine, indoramin, phenoxybenzamine, prazosin, terazosin, andtolazoline; mixed alpha+beta blockers such as bucindolol, carvedilol,and labetalol; the calcium channel blockers include dihydropyridinessuch as amlodipine, felodipine, isradipine, lercanidipine, nicardipine,nifedipine, nimodipine, and nitrendipine; non-dihydropyridines such asdiltiazem and verapamil; renin inhibitors such as Aliskiren; an ACEinhibitors such as captopril, enalapril, fosinopril, lisinopril,perindopril, quinapril, ramipril, trandolapril, and benazepril;angiotensin II receptor antagonists such as candesartan, eprosartan,irbesartan, losartan, olmesartan, telmisartan, and valsartan;aldosterone antagonists such as eplerenone and spironolactone;vasodilators such as sodium nitroprusside; and Alpha-2 agonists such asClonidine, Guanabenz, Methyldopa, Moxonidine, Guanethidine or Reserpine;or a nitrate such as nitroglycerin (glyceryl trinitrate),pentaerythritol tetranitrate, isosorbide dinitrate or isosorbidemononitrate.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering at least one otheragent, wherein the combination allows for mitigation of undesirable sideeffects of at least one other agent.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the antibody or antibody fragment or composition containing andthe at least one other agent are administered concurrently.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the antibody or antibody fragment is administered before orafter the at least one other agent.

A method for lowering serum cholesterol level in a subject comprisingadministering to a subject an effective amount of at least oneanti-human PCSK9 antibody or antibody fragment according to theinvention or a composition containing.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein administering the at least one isolated anti-human PCSK9antibody or antibody fragment or composition inhibits PCSK9 binding toLDLR.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering an agent thatelevates the availability of LDLR protein or which decreases serumcholesterol.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe agent that elevates the availability of LDLR protein or whichdecreases cholesterol is administered simultaneously with the antibodyor composition containing.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent that elevates the availability of LDLR protein orwhich decrease serum cholesterol is administered sequentially.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent that reduces serum cholesterol comprises a statin.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the statin comprises atorvastatin, cerivastatin, fluvastatin,lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin,simvastatin, or a combination thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe method further comprises administering an agent that elevates theHDL level, an agent that decreases triglyceride levels, or both.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises a fibrate.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the fibrate comprises bezafibrate, ciprofibrate, clofibrate,gemfibrozil, fenofibrate, or mixtures thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe method further comprises administering a bile sequestering agent.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe bile sequestering agent comprises cholestyramine, colesevelam,colestipol, or mixtures thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering an agent thatdecreases cholesterol absorption in the intestines.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises ezetimibe.

Another preferred embodiment of the invention relates to a method of anyone containing an antibody according to the invention wherein the methodfurther comprises administering an agent that inhibits hepatictriglyceride production, inhibits VLDL secretions, or both.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises acipimox, Gemfibrozil or nicotinic acid, andnicotinic acid analogs.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering an agent thatdecreases lipid absorption in the intestines.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises orlistat, lipstatin, or mixtures thereof.

Another preferred embodiment of the invention relates to a method ofusing an effective amount of at least one antibody according to theinvention or a composition containing optionally with another activeagent wherein the antibody or antibody fragment is administered beforeor after other active agent.

Another embodiment relates to a method of increasing LDLR protein levelin a subject, the method comprising administering to a subject aneffective amount of at least one isolated antibody or antibody fragmentaccording to the invention or a composition containing as describedherein.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein administering the at least one isolated anti-human PCSK9antibody or antibody fragment inhibits PCSK9 binding to LDLR.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering another agent thatelevates the availability of LDLR protein or one which decreases serumcholesterol.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent that elevates the availability of LDLR protein orwhich decrease serum cholesterol is administered simultaneously with theantibody or antibody composition.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent that elevates the availability of LDLR protein orwhich reduces serum cholesterol is administered sequentially withrespect to the antibody or antibody composition.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent that reduces serum cholesterol protein comprises astatin.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the statin comprises atorvastatin, cerivastatin, fluvastatin,lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin,simvastatin, or a combination thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering an agent thatelevates the HDL level, an agent that decreases triglyceride levels, orboth.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the other agent comprises a fibrate.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the fibrate comprises bezafibrate, ciprofibrate, clofibrate,gemfibrozil, fenofibrate, or mixtures thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent 234,wherein the method further comprises administering a bile sequesteringagent.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe bile sequestering agent comprises cholestyramine, colesevelam,colestipol, or mixtures thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe method further comprises administering an agent that decreasescholesterol absorption in the intestines.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises ezetimibe.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering an agent thatinhibits hepatic triglyceride production, inhibits VLDL secretions, orboth.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the agent comprises acipimox, Gemfibrozil or nicotinic acid, ora nicotinic acid analog.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering an agent thatdecreases lipid absorption in the intestines.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent 250,wherein the agent comprises orlistat, lipstatin, or mixtures thereof.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the method further comprises administering an agent that is ananti-hypertensive, treats angina, or both.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe agent comprises amlodipine, diuretic, an adrenergic receptorantagonist, a calcium channel blocker, a renin inhibitor, an ACEinhibitor, an angiotensin II receptor antagonist, aa aldosteroneantagonist, a vasodilator, a nitrate or an alpha-2 agonist.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the diuretics include bumetanide, thacrynic acid, furosemide,and torsemide; a thiazide diuretic such as epitizide,hydrochlorothiazide or chlorothiazide, or bendroflumethiazide; athiazide-like diuretic such as indapamide, chlorthalidone, ormetolazone; a potassium-sparing diuretic such as amiloride, triamterene,or spironolactone; the adrenergic receptor antagonists include betablockers such as atenolol, metoprolol, nadolol, oxprenolol, pindolol,propranolol, or timolol; the alpha blockers include doxazosin,phentolamine, indoramin, phenoxybenzamine, prazosin, terazosin, andtolazoline; mixed alpha+beta blockers such as bucindolol, carvedilol,and labetalol; the calcium channel blockers include dihydropyridinessuch as amlodipine, felodipine, isradipine, lercanidipine, nicardipine,nifedipine, nimodipine, and nitrendipine; non-dihydropyridines such asdiltiazem and verapamil; renin inhibitors such as Aliskiren; an ACEinhibitors such as captopril, enalapril, fosinopril, lisinopril,perindopril, quinapril, ramipril, trandolapril, and benazepril;angiotensin II receptor antagonists such as candesartan, eprosartan,irbesartan, losartan, olmesartan, telmisartan, and valsartan;aldosterone antagonists such as eplerenone and spironolactone;vasodilators such as sodium nitroprusside; and Alpha-2 agonists such asClonidine, Guanabenz, Methyldopa, Moxonidine, Guanethidine or Reserpine;or a nitrate such as nitroglycerin (glyceryl trinitrate),pentaerythritol tetranitrate, isosorbide dinitrate or isosorbidemononitrate.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe antibody or antibody fragment or composition containing and at leastone other agent are administered concurrently.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe antibody or antibody fragment or composition containing and at leastone other agent are administered separately, optionally by differentdosing methods.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe antibody or antibody fragment is administered before or after the atleast one other agent.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent whereinthe antibody or antibody fragment or composition containing areadministered with an agent that inhibits inflammation.

Another preferred embodiment of the invention relates to a method usingan effective amount of at least one antibody according to the inventionor a composition containing optionally with another active agent,wherein the therapeutic agent for inflammation comprises acyclooxygenase type 1 inhibitor, a cyclooxygenase type 2 inhibitor, asmall molecule modulator of p38-MAPK, a small molecule modulator ofintracellular molecules involved in inflammation pathways, or mixturesthereof.

Another preferred embodiment relates to a method using an effectiveamount of at least one antibody according to the invention or acomposition containing optionally with another active agent, that isused to treat or prevent a disease or the complications of a disease orcondition selected from hypercholesterolemia, heart disease, metabolicsyndrome, diabetes, coronary heart disease, stroke, cardiovasculardiseases, Alzheimer's disease, dyslipidemias, elevated total serumcholesterol, elevated LDL, elevated triglycerides, elevated VLDL, lowHDL, metabolic syndrome, diabetes mellitus, familial combinedhyperlipidemia, familial hypertriglyceridemia, familialhypercholesterolemias, including heterozygous hypercholesterolemia,homozygous hypercholesterolemia, familial defective apoplipoproteinB-100; polygenic hypercholesterolemia; remnant removal disease, hepaticlipase deficiency; dyslipidemia secondary to any of the following:dietary indiscretion, hypothyroidism, drugs including estrogen andprogestin therapy, beta-blockers, and thiazide diuretics; nephroticsyndrome, chronic renal failure, Cushing's syndrome, primary biliarycirrhosis, glycogen storage diseases, hepatoma, cholestasis, acromegaly,insulinoma, isolated growth hormone deficiency, and alcohol-inducedhypertriglyceridemia. atherosclerotic diseases such as coronary heartdisease, coronary artery disease, peripheral arterial disease, stroke(ischaemic and hemorrhagic), angina pectoris, cerebrovascular diseaseand acute coronary syndrome, myocardial infarction, reducing the riskof: nonfatal heart attacks, fatal and non-fatal strokes, certain typesof heart surgery, hospitalization for heart failure, chest pain inpatients with heart disease, and/or cardiovascular events because ofestablished heart disease such as prior heart attack, prior heartsurgery, and/or chest pain with evidence of clogged arteries, or therisk of recurrent cardiovascular events.

Another preferred embodiment of the invention relates to an isolatednucleic acid or nucleic acids which encode for and when expressed in asuitable host cell result in the expression of an anti-human PCSK9antibody or antibody fragment or the variable heavy or light chainthereof, wherein said antibody or antibody fragment encoded by saidnucleic acid or nucleic acids competes with and/or specifically binds tothe same or overlapping epitope(s) on human PCSK9 as an anti-human PCSK9antibody selected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5,Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17,Ab18, Ab19, Ab20, Ab21, Ab22, Ab23 and Ab24.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention wherein said expressed antibody or antibody fragmentspecifically binds to the same or overlapping epitope(s) on human PCSK9as an anti-human PCSK9 antibody selected from the group consisting ofAb1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13,Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23 and Ab24.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention, wherein said epitope(s) can be identified is a bindingassay that detects the binding of an anti-human PCSK9 antibody to one ormore peptides in a library of 10-15-mer peptides which are overlappingfragments that correspond to all or substantially all of the length ofthe human PCSK9 polypeptide.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention wherein the epitope bound thereby is determined using aWestern immunoblot assay which detects specific binding of the antibodyor antibody fragment to one or more of said 10-15 mer peptides in saidlibrary by the use of a chemiluminescent label which emits a detectablechemiluminescent signal when specific binding of said antibody orantibody fragment to a peptide in said library occurs.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention, wherein said expressed antibody or antibody fragmentencoded by said nucleic acid or nucleic acids contains at least 2complementarity determining regions (CDRs) of an anti-PCSK9 antibodyselected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19,Ab20, Ab21, Ab22, Ab23 and Ab24.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention, wherein said expressed anti-human anti-PCSK9 antibodyor antibody fragment encoded by said nucleic acid or nucleic acidscontains at least 3 complementarity determining regions (CDRs) of ananti-PCSK9 antibody selected from the group consisting of Ab1, Ab2, Ab3,Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16,Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23 and Ab24.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention, wherein said expressed anti-human PCSK9 antibody orantibody fragment encoded by said nucleic acid or nucleic acids containsat least 4 complementarity determining regions (CDRs) of an anti-PCSK9antibody selected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5,Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17,Ab18, Ab19, Ab20, Ab21, Ab22, Ab23 and Ab24.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention, wherein said expressed anti-human PCSK9 antibody orantibody fragment encoded by said nucleic acid or nucleic acids containsat least 5 complementarity determining regions (CDRs) of an anti-PCSK9antibody selected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5,Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17,Ab18, Ab19, Ab20, Ab21, Ab22, Ab23 and Ab24.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention wherein said expressed antibody or antibody fragmentencoded by said nucleic acid or nucleic acids contains all 6complementarity determining regions (CDRs) of an anti-PCSK9 antibodyselected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19,Ab20, Ab21, Ab22, Ab23 and Ab24.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention wherein said expressed anti-human PCSK9 antibody orantibody fragment encoded by said nucleic acid or nucleic acids contains(a) a variable heavy chain comprising a CDR1 sequence selected from thegroup consisting of SEQ ID NO: 4, SEQ ID NO: 44, SEQ ID NO: 84, SEQ IDNO: 124, SEQ ID NO: 164, SEQ ID NO: 204, SEQ ID NO: 244, SEQ ID NO: 284,SEQ ID NO: 324, SEQ ID NO: 364, SEQ ID NO: 404, SEQ ID NO: 444, SEQ IDNO: 484, SEQ ID NO: 524, SEQ ID NO: 564, SEQ ID NO: 604, SEQ ID NO: 644,SEQ ID NO: 684, SEQ ID NO: 724, SEQ ID NO: 764, SEQ ID NO: 804, SEQ IDNO: 844, SEQ ID NO: 884, and SEQ ID NO: 924; a CDR2 sequence selectedfrom the group consisting of SEQ ID NO: 6, SEQ ID NO: 46, SEQ ID NO: 86,SEQ ID NO: 126, SEQ ID NO: 166, SEQ ID NO: 206, SEQ ID NO: 246, SEQ IDNO: 286, SEQ ID NO: 326, SEQ ID NO: 366, SEQ ID NO: 406, SEQ ID NO: 446,SEQ ID NO: 486, SEQ ID NO: 526, SEQ ID NO: 566, SEQ ID NO: 606, SEQ IDNO: 646, SEQ ID NO: 686, SEQ ID NO: 726, SEQ ID NO: 766, SEQ ID NO: 806,SEQ ID NO: 846, SEQ ID NO: 886, and SEQ ID NO: 926; and a CDR3 sequenceselected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 48, SEQID NO: 88, SEQ ID NO: 128, SEQ ID NO: 168, SEQ ID NO: 208, SEQ ID NO:248, SEQ ID NO: 288, SEQ ID NO: 328, SEQ ID NO: 368, SEQ ID NO: 408, SEQID NO: 448, SEQ ID NO: 488, SEQ ID NO: 528, SEQ ID NO: 568, SEQ ID NO:608, SEQ ID NO: 648, SEQ ID NO: 688, SEQ ID NO: 728, SEQ ID NO: 768, SEQID NO: 808, SEQ ID NO: 848, SEQ ID NO: 888, and SEQ ID NO: 928; and/or

-   -   (b) a variable light chain comprising a CDR1 sequence selected        from the group consisting of SEQ ID NO: 24, SEQ ID NO: 64, SEQ        ID NO: 104, SEQ ID NO: 144, SEQ ID NO: 184, SEQ ID NO: 224, SEQ        ID NO: 264, SEQ ID NO: 304, SEQ ID NO: 344, SEQ ID NO: 384, SEQ        ID NO: 424, SEQ ID NO: 464, SEQ ID NO: 504, SEQ ID NO: 544, SEQ        ID NO: 584, SEQ ID NO: 624, SEQ ID NO: 664, SEQ ID NO: 704, SEQ        ID NO: 744, SEQ ID NO: 784, SEQ ID NO: 824, SEQ ID NO: 864, SEQ        ID NO: 904, and SEQ ID NO: 944; a CDR2 sequence selected from        the group consisting of SEQ ID NO: 26, SEQ ID NO: 66, SEQ ID NO:        106, SEQ ID NO: 146, SEQ ID NO: 186, SEQ ID NO: 226, SEQ ID NO:        266, SEQ ID NO: 306, SEQ ID NO: 346, SEQ ID NO: 386, SEQ ID NO:        426, SEQ ID NO: 466, SEQ ID NO: 506, SEQ ID NO: 546, SEQ ID NO:        586, SEQ ID NO: 626, SEQ ID NO: 666, SEQ ID NO: 706, SEQ ID NO:        746, SEQ ID NO: 786, SEQ ID NO: 826, SEQ ID NO: 866, SEQ ID NO:        906, and SEQ ID NO: 946; and a CDR3 sequence selected from the        group consisting of SEQ ID NO: 28, SEQ ID NO: 68, SEQ ID NO:        108, SEQ ID NO: 148, SEQ ID NO: 188, SEQ ID NO: 228, SEQ ID NO:        268, SEQ ID NO: 308, SEQ ID NO: 348, SEQ ID NO: 388, SEQ ID NO:        428, SEQ ID NO: 468, SEQ ID NO: 508, SEQ ID NO: 548, SEQ ID NO:        588, SEQ ID NO: 628, SEQ ID NO: 668, SEQ ID NO: 708, SEQ ID NO:        748, SEQ ID NO: 788, SEQ ID NO: 828, SEQ ID NO: 868, SEQ ID NO:        908, and SEQ ID NO: 948, with the further proviso that one or        two residues of any of the afore-identified CDR polypeptides may        be substituted with another amino acid.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention, wherein the variable heavy chain encoded by saidnucleic acid or nucleic acids comprises the CDR1 sequence of SEQ ID NO:4, the CDR2 sequence of SEQ ID NO: 6, and the CDR3 sequence of SEQ IDNO: 8; and/or the variable light chain comprises the CDR1 sequence ofSEQ ID NO: 24, the CDR2 sequence of SEQ ID NO: 26, and the CDR3 sequenceof SEQ ID NO: 28.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention, wherein the variable heavy chain encoded by saidnucleic acid or nucleic acids comprises the CDR1 sequence of SEQ ID NO:44, the CDR2 sequence of SEQ ID NO: 46, and the CDR3 sequence of SEQ IDNO: 48; and/or the variable light chain comprises the CDR1 sequence ofSEQ ID NO: 64, the CDR2 sequence of SEQ ID NO: 66, and the CDR3 sequenceof SEQ ID NO: 68.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that express an antibody or antibody fragment accordingto the invention wherein the variable heavy chain encoded by saidnucleic acid or nucleic acids comprises the CDR1 sequence of SEQ ID NO:84, the CDR2 sequence of SEQ ID NO: 86, and the CDR3 sequence of SEQ IDNO: 88; and/or the variable light chain comprises the CDR1 sequence ofSEQ ID NO: 104, the CDR2 sequence of SEQ ID NO: 106, and the CDR3sequence of SEQ ID NO: 108.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 124, the CDR2 sequenceof SEQ ID NO: 126, and the CDR3 sequence of SEQ ID NO: 128; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 144, theCDR2 sequence of SEQ ID NO: 146, and the CDR3 sequence of SEQ ID NO:148.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 164, the CDR2 sequenceof SEQ ID NO: 166, and the CDR3 sequence of SEQ ID NO: 168; and thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 184, theCDR2 sequence of SEQ ID NO: 186, and the CDR3 sequence of SEQ ID NO:188.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein said expressed anti-human PCSK9 antibody or antibody fragmentcontains a variable heavy chain that comprises the CDR1 sequence of SEQID NO: 204, the CDR2 sequence of SEQ ID NO: 206, and the CDR3 sequenceof SEQ ID NO: 208; and the variable light chain comprises the CDR1sequence of SEQ ID NO: 224, the CDR2 sequence of SEQ ID NO: 226, and theCDR3 sequence of SEQ ID NO: 228.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 244, the CDR2 sequenceof SEQ ID NO: 246, and the CDR3 sequence of SEQ ID NO: 248; and thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 264, theCDR2 sequence of SEQ ID NO: 266, and the CDR3 sequence of SEQ ID NO:268.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 284, the CDR2 sequenceof SEQ ID NO: 286, and the CDR3 sequence of SEQ ID NO: 288; and thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 304, theCDR2 sequence of SEQ ID NO: 306, and the CDR3 sequence of SEQ ID NO:308.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 324, the CDR2 sequenceof SEQ ID NO: 326, and the CDR3 sequence of SEQ ID NO: 328; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 344, theCDR2 sequence of SEQ ID NO: 346, and the CDR3 sequence of SEQ ID NO:348.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 364, the CDR2 sequenceof SEQ ID NO: 366, and the CDR3 sequence of SEQ ID NO: 368; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 384, theCDR2 sequence of SEQ ID NO: 386, and the CDR3 sequence of SEQ ID NO:388.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 404, the CDR2 sequenceof SEQ ID NO: 406, and the CDR3 sequence of SEQ ID NO: 408; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 424, theCDR2 sequence of SEQ ID NO: 426, and the CDR3 sequence of SEQ ID NO:428.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 444, the CDR2 sequenceof SEQ ID NO: 446, and the CDR3 sequence of SEQ ID NO: 448; and thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 464, theCDR2 sequence of SEQ ID NO: 466, and the CDR3 sequence of SEQ ID NO:468.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 484, the CDR2 sequenceof SEQ ID NO: 486, and the CDR3 sequence of SEQ ID NO: 488; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 504, theCDR2 sequence of SEQ ID NO: 506, and the CDR3 sequence of SEQ ID NO:508.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 524, the CDR2 sequenceof SEQ ID NO: 526, and the CDR3 sequence of SEQ ID NO: 528; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 544, theCDR2 sequence of SEQ ID NO: 546, and the CDR3 sequence of SEQ ID NO:548.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 564, the CDR2 sequenceof SEQ ID NO: 566, and the CDR3 sequence of SEQ ID NO: 568; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 584, theCDR2 sequence of SEQ ID NO: 586, and the CDR3 sequence of SEQ ID NO:588.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 604, the CDR2 sequenceof SEQ ID NO: 606, and the CDR3 sequence of SEQ ID NO: 608; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 624, theCDR2 sequence of SEQ ID NO: 626, and the CDR3 sequence of SEQ ID NO:628.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 644, the CDR2 sequenceof SEQ ID NO: 646, and the CDR3 sequence of SEQ ID NO: 648; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 664, theCDR2 sequence of SEQ ID NO: 666, and the CDR3 sequence of SEQ ID NO:668.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 684, the CDR2 sequenceof SEQ ID NO: 686, and the CDR3 sequence of SEQ ID NO: 688; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 704, theCDR2 sequence of SEQ ID NO: 706, and the CDR3 sequence of SEQ ID NO:708.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 724, the CDR2 sequenceof SEQ ID NO: 726, and the CDR3 sequence of SEQ ID NO: 728; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 744, theCDR2 sequence of SEQ ID NO: 746, and the CDR3 sequence of SEQ ID NO:748.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 764, the CDR2 sequenceof SEQ ID NO: 766, and the CDR3 sequence of SEQ ID NO: 768; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 784, theCDR2 sequence of SEQ ID NO: 786, and the CDR3 sequence of SEQ ID NO:788.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 804, the CDR2 sequenceof SEQ ID NO: 806, and the CDR3 sequence of SEQ ID NO: 808; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 824, theCDR2 sequence of SEQ ID NO: 826, and the CDR3 sequence of SEQ ID NO:828.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 844, the CDR2 sequenceof SEQ ID NO: 846, and the CDR3 sequence of SEQ ID NO: 848; and/or thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 864, theCDR2 sequence of SEQ ID NO: 866, and the CDR3 sequence of SEQ ID NO:868.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 884, the CDR2 sequenceof SEQ ID NO: 886, and the CDR3 sequence of SEQ ID NO: 888; and/or thevariable light chain encoded by said nucleic acid or nucleic acidscomprises the CDR1 sequence of SEQ ID NO: 904, the CDR2 sequence of SEQID NO: 906, and the CDR3 sequence of SEQ ID NO: 908.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises the CDR1 sequence of SEQ ID NO: 924, the CDR2 sequenceof SEQ ID NO: 926, and the CDR3 sequence of SEQ ID NO: 928; and/or thevariable light chain encoded by said nucleic acid or nucleic acidscomprises the CDR1 sequence of SEQ ID NO: 944, the CDR2 sequence of SEQID NO: 946, and the CDR3 sequence of SEQ ID NO: 948.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 2 and/or the variable light chain encodedby said nucleic acid or nucleic acids comprises SEQ ID NO: 22.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 2 and SEQID NO: 22, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 42 and the variable light chain encoded bysaid nucleic acid or nucleic acids comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 62.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 42 andSEQ ID NO: 62, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 82 and the variable light chain encoded bysaid nucleic acid or nucleic acids comprises a sequence at least 80, 85,90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 102.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 82 andSEQ ID NO: 102, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 122 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 142.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 122 andSEQ ID NO: 142, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 162 and the variable light chain encoded bysaid nucleic acid or nucleic acids comprises encoded by said nucleicacid or nucleic acids comprises a sequence at least 80, 85, 90, 95, 96,97, 98, 99 or 100% identical to SEQ ID NO: 182.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 162 andSEQ ID NO: 182, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 202 and the variable light chain encoded bysaid nucleic acid or nucleic acids comprises encoded by said nucleicacid or nucleic acids comprises a sequence at least 80, 85, 90, 95, 96,97, 98, 99 or 100% identical to SEQ ID NO: 222.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 202 andSEQ ID NO: 222, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 242 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 262.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention, 47,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 242 andSEQ ID NO: 262, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 282 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 302.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 282 andSEQ ID NO: 302, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 322 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 342.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 322 andSEQ ID NO: 342, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 362 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 382.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 362 andSEQ ID NO: 382, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 402 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 422.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention, 55,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 402 andSEQ ID NO: 422, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 442 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 462.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 442 andSEQ ID NO: 462, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 482 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 502.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 482 andSEQ ID NO: 502, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 522 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 542.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 522 andSEQ ID NO: 542, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 562 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 582.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 562 andSEQ ID NO: 582, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 602 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 622.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 602 andSEQ ID NO: 622, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 642 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 662.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 642 andSEQ ID NO: 662, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 682 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 702.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 682 andSEQ ID NO: 702, respectively

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 722 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 742.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 722 andSEQ ID NO: 742, respectively

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 762 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 782.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 762 andSEQ ID NO: 782, respectively

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 802 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 822.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 802 andSEQ ID NO: 822, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 842 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 862.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 842 andSEQ ID NO: 862, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 882 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 902.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 882 andSEQ ID NO: 902, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 922 and the variable light chain encoded by said nucleic acidor nucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 942.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the variable heavy and light chains are each at least 90%identical to the variable heavy and light chains in SEQ ID NO: 922 andSEQ ID NO: 942, respectively.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises encoded by said nucleic acid or nucleic acids comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 1 and the light chain encoded by said nucleic acid or nucleicacids comprises encoded by said nucleic acid or nucleic acids comprisesa sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 21.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 41 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 61.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises encoded by said nucleic acid or nucleic acids comprises asequence is at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 81 and the light chain encoded by said nucleic acid ornucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 101.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 121 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 141.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acids asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 161 and the light chain encoded by said nucleic acid ornucleic acids comprises encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 181.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 201 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 221.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 241 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 261.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises encoded by said nucleic acid or nucleic acids comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 281 and the light chain encoded by said nucleic acid ornucleic acids comprises SEQ ID NO: 301.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises SEQ ID NO: 321 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 341.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 361 and the light chain encoded by said nucleicacid or nucleic acids comprises SEQ ID NO: 381.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises SEQ ID NO: 401 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 421.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 441 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 461.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 481 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 501.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 521 and the light chain encoded by said nucleicacid or nucleic acids comprises SEQ ID NO: 541.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises SEQ ID NO: 561 and the light chain encoded by said nucleicacid or nucleic acids comprises a sequence at least 80, 85, 90, 95, 96,97, 98, 99 or 100% identical to SEQ ID NO: 581.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acids asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 601 and the light chain encoded by said nucleic acid ornucleic acids comprises SEQ ID NO: 621.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises SEQ ID NO: 641 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 661.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises s a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100%identical to SEQ ID NO: 681 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 701.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises encoded by said nucleic acid or nucleic acids comprises asequence at least 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical toSEQ ID NO: 721 and the light chain encoded by said nucleic acid ornucleic acids comprises SEQ ID NO: 741.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises SEQ ID NO: 761 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 781.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence that is at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 801 and the light chain encoded by saidnucleic acid or nucleic acids comprises encoded by said nucleic acid ornucleic acids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98,99 or 100% identical to SEQ ID NO: 821.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence that is at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 841 and the light chain encoded by saidnucleic acid or nucleic acids comprises encoded by said nucleic acid ornucleic acids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98,99 or 100% identical to SEQ ID NO: 861.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises a sequence that is at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 881 and the light chain encoded by saidnucleic acid or nucleic acids comprises SEQ ID NO: 901.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids that encode an antibody according to the inventionwherein the heavy chain encoded by said nucleic acid or nucleic acidscomprises SEQ ID NO: 921 and the light chain encoded by said nucleicacid or nucleic acids comprises encoded by said nucleic acid or nucleicacids comprises a sequence at least 80, 85, 90, 95, 96, 97, 98, 99 or100% identical to SEQ ID NO: 941.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention whereinthe antibody or antibody fragment encoded by said nucleic acid ornucleic acids is selected from the group consisting of chimeric,humanized, and human antibodies or antibody fragments.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the expressed antibody or antibody fragment encoded by saidnucleic acid or nucleic acids is selected from the group consisting ofscFvs, camelbodies, nanobodies, IgNAR, F_(ab) fragments, F_(ab′)fragments, MetMabs, monovalent antibody fragments, and F_((ab′)2)fragments.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the expressed antibody or antibody fragment encoded by saidnucleic acid or nucleic acids substantially or entirely lacksN-glycosylation and/or O-glycosylation.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the expressed antibody or antibody fragment encoded by saidnucleic acid or nucleic acids comprises a human constant domain.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein the expressed antibody encoded by said nucleic acid or nucleicacids is an IgG1, IgG2, IgG3, or IgG4 antibody.

Another preferred embodiment of the invention relates to a nucleic acidor nucleic acids encoding an antibody according to the invention,wherein said nucleic acid or nucleic acids which encode said antibody orantibody fragment comprise a sequence encoding a VH and a VL region atleast 80, 90, 95, 96, 97, 98, 99 or 100% identical to SEQ ID NO: 12, SEQID NO: 32; SEQ ID NO: 52, SEQ ID NO: 72; SEQ ID NO: 92, SEQ ID NO: 112;SEQ ID NO: 132, SEQ ID NO: 152; SEQ ID NO: 172, SEQ ID NO: 192; SEQ IDNO: 212, SEQ ID NO: 232, SEQ ID NO: 252, SEQ ID NO: 272; SEQ ID NO: 292,SEQ ID NO: 312; SEQ ID NO: 332, SEQ ID NO: 352; SEQ ID NO: 372, SEQ IDNO: 392; SEQ ID NO: 412, SEQ ID NO: 432; SEQ ID NO: 452, SEQ ID NO: 472;SEQ ID NO: 492, SEQ ID NO: 512, SEQ ID NO: 532, SEQ ID NO: 552, SEQ IDNO: 572, SEQ ID NO: 592, SEQ ID NO: 612, SEQ ID NO: 632, SEQ ID NO: 652,SEQ ID NO: 672; SEQ ID NO: 692; SEQ ID NO: 712; SEQ ID NO: 732; SEQ IDNO: 752; SEQ ID NO: 772; SEQ ID NO: 792; SEQ ID NO: 812; SEQ ID NO: 832;SEQ ID NO: 852; SEQ ID NO: 872; SEQ ID NO: 892; SEQ ID NO: 912; SEQ IDNO: 932; SEQ ID NO: 952; or a codon degenerate thereof.

A vector or vectors comprising the nucleic acid sequence or sequencesencoding an antibody according to the invention.

A host cell comprising nucleic acid sequence or sequences encoding anantibody according to the invention or a vector or vectors containing.

Another preferred embodiment of the invention relates to an host cellcontaining a nucleic acid or acids encoding an antibody according to theinvention, wherein said host cell is a mammalian, bacterial, fungal,yeast, avian or insect cell.

Another preferred embodiment of the invention relates to an host cellencoding an antibody according to the invention, which comprises afilamentous fungi or a yeast.

Another preferred embodiment of the invention relates to an host cellencoding an antibody according to the invention, which comprises afilamentous fungi or yeast include Pichia pastoris, Pichia finlandica,Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens, Pichiaminuta (Ogataea minuta, Pichia lindneri), Pichia opuntiae, Pichiathermotolerans, Pichia salictaria, Pichia guercuum, Pichia pijperi,Pichia stiptis, Pichia methanolica, Pichia sp., Saccharomycescerevisiae, Saccharomyces sp., Hansenula polymorpha, Kluyveromyces sp.,Kluyveromyces lactis, Candida albicans, Aspergillus nidulans,Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporiumlucknowense, Fusarium sp., Fusarium gramineum, Fusarium venenatum,Physcomitrella patens and Neurospora crassa. Pichia sp., anySaccharomyces sp., Hansenula polymorpha, any Kluyveromyces sp., Candidaalbicans, any Aspergillus sp., Trichoderma reesei, Chrysosporiumlucknowense, any Fusarium sp. and Neurospora crassa.

Another preferred embodiment of the invention relates to an host cellencoding an antibody according to the invention, wherein said host cellis a mammalian cell.

Another preferred embodiment of the invention relates to an host cellencoding an antibody according to the invention, wherein said host cellis a CHO, COS, BHK, myeloma, monkey kidney CV1 line transformed by SV40(COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cellssubcloned for growth in suspension culture), mouse sertoli cell; humanlung cell; human liver cell, or a mouse mammary tumor cell.

Another preferred embodiment of the invention relates to an host cellencoding an antibody according to the invention, wherein the mammaliancell is a CHO cell.

Another preferred embodiment of the invention relates to an host cellencoding an antibody according to the invention, which is yeast cellbelonging to the genus Pichia.

Another preferred embodiment of the invention relates to an host cellencoding an antibody according to the invention, which is a Pichiapastoris.

A method of making the antibody or antibody fragment according to theinvention, by expressing nucleic acids which encode for the expressionof said antibody or antibody fragment in a recombinant host cell.

Another preferred embodiment of the invention relates to a method usinga cell that expresses an antibody according to the invention wherein thehost cell is selected from a bacteria, yeast, fungi, insect cell, plantcell, avian cell, or mammalian cell.

Another preferred embodiment of the invention relates to a method ofusing a cell that expresses an antibody according to the invention,wherein the host cell is a yeast or a filamentous fungi, which mayoptimally be polyploidal.

Another preferred embodiment of the invention relates to a method ofusing a cell that expresses an antibody according to the invention,wherein said a yeast or a filamentous fungi include Pichia pastoris,Pichia finlandica, Pichia trehalophila, Pichia koclamae, Pichiamembranaefaciens, Pichia minuta (Ogataea minuta, Pichia lindneri),Pichia opuntiae, Pichia thermotolerans, Pichia salictaria, Pichiaguercuum, Pichia pijperi, Pichia stiptis, Pichia methanolica, Pichiasp., Saccharomyces cerevisiae, Saccharomyces sp., Hansenula polymorpha,Kluyveromyces sp., Kluyveromyces lactis, Candida albicans, Aspergillusnidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei,Chrysosporium lucknowense, Fusarium sp., Fusarium gramineum, Fusariumvenenatum, Physcomitrella patens and Neurospora crassa. Pichia sp., anySaccharomyces sp., Hansenula polymorpha, any Kluyveromyces sp., Candidaalbicans, any Aspergillus sp., Trichoderma reesei, Chrysosporiumlucknowense, any Fusarium sp. and Neurospora crassa.

Another preferred embodiment of the invention relates to a method ofusing a cell that expresses an antibody according to the invention,wherein said host cell is a mammalian cell.

Another preferred embodiment of the invention relates to a method ofusing a cell that expresses an antibody according to the invention,wherein said host cell is a CHO, COS, BHK, myeloma, monkey kidney CV1line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidneyline (293 or 293 cells subcloned for growth in suspension culture),mouse sertoli cell; human lung cell; human liver cell, or a mousemammary tumor cell.

Another preferred embodiment of the invention relates to a method ofusing a cell that expresses an antibody according to the invention,wherein the mammalian cell is a CHO cell.

Another preferred embodiment of the invention relates to a method of383, wherein the recombinant host cell is a polyploid yeast culture thatstably expresses and secretes into the culture medium at least 10-25mg/liter of said antibody or antibody fragment.

Another preferred embodiment of the invention relates to a method ofusing a cell that expresses an antibody according to the invention,wherein the cell is a polyploidal yeast made by a method that comprises:

-   -   (i) introducing at least one expression vector containing one or        more heterologous polynucleotides encoding said antibody        operably linked to a promoter and a signal sequence into a        haploid yeast cell;    -   (ii) producing by mating or spheroplast fusion a polyploidal        yeast from said first and/or second haploid yeast cell;    -   (iii) selecting polyploidal yeast cells that stably express said        antibody; and    -   (iv) producing stable polyploidal yeast cultures from said        polyploidal yeast cells that stably express said antibody into        the culture medium.

Another preferred embodiment of the invention relates to a method ofusing a yeast cell that expresses an antibody according to theinvention, wherein said yeast is of the genera Pichia.

Another preferred embodiment relates to any therapeutic or diagnosticmethod that comprises the administration of a therapeutically ordiagnostically effective amount of at least one antibody or antibodyfragment according to the invention.

Another preferred embodiment of the invention relates to a method ofusing an antibody according to the invention, wherein the antibody orantibody fragment is administered by a means selected from buccal,epicutaneous, epidural, inhalation, intraarterial, intracardial,intracerebroventricular, intradermal, intramuscular, intranasal,intraocular, intraperitoneal, intraspinal, intrathecal, intravenous,oral, parenteral, rectally via an enema or suppository, subcutaneous,subdermal, sublingual, transdermal, and transmucosal.

Another preferred embodiment of the invention relates to a method ofusing an antibody according to the invention, wherein the antibody orantibody fragment is administered by a means selected from subcutaneous,intravenous, intraarterial, intracardial, intracerebroventricular,intramuscular, intraperitoneal, intraspinal, intrathecal, andparenteral.

Another preferred embodiment of the invention relates to a method ofusing an antibody according to the invention, wherein the antibody orantibody fragment is administered by subcutaneous means.

Another preferred embodiment of the invention relates to a method ofusing an antibody according to the invention wherein the antibody orantibody fragment is administered by intravenous means.

Another preferred embodiment of the invention relates to a method ofusing an antibody according to the invention, wherein the antibody orantibody fragment is contained in a powder, liquid, gel, drop, or othermeans of administration.

Another preferred embodiment of the invention relates to a compositioncontaining an antibody or antibody fragment according to the invention,which is suitable for administration by a means selected from buccal,epicutaneous, epidural, inhalation, intraarterial, intracardial,intracerebroventricular, intradermal, intramuscular, intranasal,intraocular, intraperitoneal, intraspinal, intrathecal, intravenous,oral, parenteral, rectally via an enema or suppository, subcutaneous,subdermal, sublingual, transdermal, and transmucosal.

Another preferred embodiment of the invention relates to an compositioncontaining an antibody according to the invention, wherein thecomposition is administrable by a means selected from subcutaneous,intravenous, intraarterial, intracardial, intracerebroventricular,intramuscular, intraperitoneal, intraspinal, intrathecal, andparenteral.

Another preferred embodiment of the invention relates to an compositionencoding an antibody according to the invention, which is administrableby subcutaneous means.

Another preferred embodiment of the invention relates to a compositionwhich is administrable by intravenous means.

Another preferred embodiment of the invention relates to a compositionwhich comprises a powder, liquid, gel, drop, liposomal, or other dosageform.

Another preferred embodiment of the invention relates to a method ofusing an antibody or antibody fragment according to the invention toisolate, detect or purify PCSK9 in a sample.

Another preferred embodiment of the invention relates a method of usingan antibody or antibody fragment according to the invention to detectthe levels of PCSK9 in vivo or ex vivo.

Another preferred embodiment of the invention relates to a method ofusing the subject antibodies to assess whether a patient should beadministered an anti-PCSK9 antibody or antibody fragment or othertherapy.

Another preferred embodiment of the invention relates to detectionmethod using the subject antibodies to assess the efficacy of atreatment method the objective of which is to reduce cholesterol oralter lipid levels.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

Antibodies and binding fragments thereof that bind to PCSK9 aredisclosed herein. In some embodiments, the antibody or antibody fragmentaccording to the invention bind to PCSK9 and prevent PCSK9 fromfunctioning in various ways. In some embodiments, the antibody orantibody fragment according to the invention block or reduce the abilityof PCSK9 to interact with other substances, e.g., LDLR. For example, insome embodiments, the antibody or antibody fragment according to theinvention binds to PCSK9 in a manner that prevents or reduces thelikelihood that PCSK9 will bind to LDLR. In other embodiments, antibodyor antibody fragment according to the invention bind to PCSK9 but do notblock PCSK9's ability to interact with LDLR. In some embodiments, theantibody or antibody fragment according to the invention are humanmonoclonal antibodies.

As will be appreciated by one of skill in the art, altering theinteractions between PCSK9 and LDLR can increase the amount of LDLRavailable for binding to LDL, which in turn decreases the amount ofserum LDL-C in a subject, resulting in a reduction in the subject'sserum cholesterol level. As such, an antibody or antibody fragmentaccording to the invention can be used in various methods andcompositions for treating subjects with elevated serum cholesterollevels, at risk of elevated serum cholesterol levels, or which couldbenefit from a reduction in their serum cholesterol levels. Theseconditions include any of the afore-mentioned conditions. Thus, variousmethods and techniques for lowering, maintaining, or preventing anincrease in serum cholesterol are also described herein. In someembodiments, the antibody or antibody fragment according to theinvention allows for binding between PCSK9 and LDLR, but the antibody orantibody fragment according to the invention prevents or reduces theadverse activity of PCSK9 on LDLR. In some embodiments, the antibody orantibody fragment according to the invention prevents or reduces thebinding of PCSK9 to LDLR.

For convenience, the following sections generally outline the variousmeanings of the terms used herein. Following this discussion, generalaspects regarding antibody or antibody fragment according to theinvention are discussed, followed by specific examples demonstrating theproperties of various embodiments of the antibody or antibody fragmentaccording to the invention and how they can be employed.

DEFINITIONS

It is to be understood that this invention is not limited to theparticular methodology, protocols, cell lines, animal species or genera,and reagents described, as such may vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to limit the scope ofthe present invention which will be limited only by the appended claims.As used herein the singular forms “a”, “and”, and “the” include pluralreferents unless the context clearly dictates otherwise. Thus, forexample, reference to “a cell” includes a plurality of such cells andreference to “the protein” includes reference to one or more proteinsand equivalents thereof known to those skilled in the art, and so forth.All technical and scientific terms used herein have the same meaning ascommonly understood to one of ordinary skill in the art to which thisinvention belongs unless clearly indicated otherwise.

The term “proprotein convertase subtilisin kexin type 9” or “PCSK9”refers to a polypeptide as set forth in SEQ ID NO: 961 and/or SEQ ID NO:962 or fragments thereof, as well as related polypeptides, whichinclude, but are not limited to, allelic variants, splice variants,derivative variants, substitution variants, deletion variants, and/orinsertion variants including the addition of an N-terminal methionine,fusion polypeptides, and interspecies homologs. In certain embodiments,a PCSK9 polypeptide includes terminal residues, such as, but not limitedto, leader sequence residues, targeting residues, amino terminalmethionine residues, lysine residues, tag residues, and/or fusionprotein residues. “PCSK9” has also been referred to as FH3, NARC1,HCHOLA3, proprotein convertase subtilisin/kexin type 9, and neuralapoptosis regulated convertase 1. The PCSK9 gene encodes a proproteinconvertase protein that belongs to the proteinase K subfamily of thesecretory subtilase family. The term “PCSK9” denotes both the proprotein(SEQ ID NO: 961) and the product generated following autocatalysis ofthe proprotein (SEQ ID NO: 962). When only the autocatalyzed product isbeing referred to (such as for an antibody or antibody fragmentaccording to the invention that selectively binds to the cleaved PCSK9),the protein can be referred to as the “mature,” “cleaved”, “processed”or “active” PCSK9. When only the inactive form is being referred to, theprotein can be referred to as the “inactive”, “pro-form”, or“unprocessed” form of PCSK9. The term PCSK9 as used herein also includesnaturally occurring alleles, such as the mutations D374Y, S127R andF216L. The term PCSK9 also encompasses PCSK9 molecules incorporatingpost-translational modifications of the PCSK9 amino acid sequence, suchas PCSK9 sequences that have been glycosylated, PEGylated, PCSK9sequences from which its signal sequence has been cleaved, PCSK9sequence from which its pro domain has been cleaved from the catalyticdomain but not separated from the catalytic domain.

The term “PCSK9 activity” includes any biological effect of PCSK9. Incertain embodiments, PCSK9 activity includes the ability of PCSK9 tointeract or bind to a substrate or receptor. In some embodiments, PCSK9activity is represented by the ability of PCSK9 to bind to a LDL-Rreceptor (LDLR). In some embodiments, PCSK9 binds to and catalyzes areaction involving LDLR. In some embodiments, PCSK9 activity includesthe ability of PCSK9 to alter (e.g., reduce) the availability of LDLR.In some embodiments, PCSK9 activity includes the ability of PCSK9 toincrease the amount of LDL-C in a subject. In some embodiments, PCSK9activity includes the ability of PCSK9 to decrease the amount of LDLRthat is available to bind to LDL. In some embodiments, “PCSK9 activity”includes any biological activity resulting from PCSK9 signaling.Exemplary activities include, but are not limited to, PCSK9 binding toLDLR, PCSK9 enzyme activity that cleaves LDLR or other proteins, PCSK9binding to proteins other than LDLR that facilitate PCSK9 action, PCSK9altering APOB secretion (Sun X-M et al, “Evidence for effect of mutantPCSK9 on apoliprotein B secretion as the cause of unusually severedominant hypercholesterolemia, Human Molecular Genetics 14: 1161-1169,2005 and Ouguerram K et al, “Apolipoprotein B100 metabolism inautosomal-dominant hypercholesterolemia related to mutations in PCSK9,Arterioscler Thromb Vasc Biol. 24: 1448-1453, 2004), PCSK9's role inliver regeneration and neuronal cell differentiation (Seidah N G et al,“The secretory proprotein convertase neural apoptosis-regulatedconvertase 1 (NARC-1): Liver regeneration and neuronal differentiation”PNAS 100: 928-933, 2003), and PCSK9's role in hepatic glucose metabolism(Costet et al., “Hepatic PCSK9 expression is regulated by nutritionalstatus via insulin and sterol regulatory element-binding protein 1c” J.Biol. Chem. 281(10):6211-18, 2006).

The term “hypercholesterolemia,” as used herein, refers to a conditionin which cholesterol levels are elevated above a desired level. In someembodiments, this denotes that serum cholesterol levels are elevated. Insome embodiments, the desired level takes into account various “riskfactors” that are known to one of skill in the art (and are described orreferenced herein).

The term “Recombinant cell” or “recombinant host cell” herein in generalrefers to any cell engineered to express one or more antibodypolypeptides according to the invention. This includes by way of examplebacterial, fungal, yeast, mammalian, invertebrate such as insect, plantand avian cells. Preferred host cells are yeast, fungi, especiallyfilamentous fungi and mammalian cells. Yeast and filamentous fungiinclude, but are not limited to Pichia pastoris, Pichia finlandica,Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens, Pichiaminuta (Ogataea minuta, Pichia lindneri), Pichia opuntiae, Pichiathermotolerans, Pichia salictaria, Pichia guercuum, Pichia pijperi,Pichia stiptis, Pichia methanolica, Pichia sp., Saccharomycescerevisiae, Saccharomyces sp., Hansenula polymorpha, Kluyveromyces sp.,Kluyveromyces lactis, Candida albicans, Aspergillus nidulans,Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporiumlucknowense, Fusarium sp., Fusarium gramineum, Fusarium venenatum,Physcomitrella patens and Neurospora crassa. Pichia sp., anySaccharomyces sp., Hansenula polymorpha, any Kluyveromyces sp., Candidaalbicans, any Aspergillus sp., Trichoderma reesei, Chrysosporiumlucknowense, any Fusarium sp. and Neurospora crassa.

Examples of invertebrate cells include insect cells such as DrosophilaS2 and Spodoptera Sf9, as well as plant cells. Examples of usefulmammalian host cell lines include Chinese hamster ovary (CHO) and COScells. More specific examples include monkey kidney CV1 line transformedby SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293cells subcloned for growth in suspension culture, Graham et al., J. GenVirol., 36:59 (1977)); Chinese hamster ovary cells/−DHFR(CHO, Urlaub andChasin, Proc. Natl. Acad. Sci. USA, 77:4216 (1980)); mouse sertoli cells(TM4, Mather, Biol. Reprod., 23:243-251 (1980)); human lung cells (W138,ATCC CCL 75); human liver cells (Hep G2, HB 8065); and mouse mammarytumor (MMT 060562, ATCC CCL51). The selection of the appropriate hostcell is deemed to be within the skill in the art. Preferred mammaliancells for antibody expression include CHO cells and COS cells. In anexemplary embodiment the recombinant host cells are polyploidal yeastcells of the genus Pichia.

Mating Competent Yeast Species:

In the present invention this is intended to broadly encompass anydiploid or tetraploid yeast which can be grown in culture. Such speciesof yeast may exist in a haploid, diploid, or other polyploid form. Thecells of a given ploidy may, under appropriate conditions, proliferatefor an indefinite number of generations in that form. Diploid cells canalso sporulate to form haploid cells. Sequential mating can result intetraploid strains through further mating or fusion of diploid strains.The present invention contemplates the use of haploid yeast, as well asdiploid or other polyploid yeast cells produced, for example, by matingor spheroplast fusion.

Mating competent yeast include yeast which are a member of theSaccharomycetaceae family, which includes the genera Arxiozyma;Ascobotryozyma; Citeromyces; Debaryomyces; Dekkera; Eremothecium;Issatchenkia; Kazachstania; Kluyveromyces; Kodamaea; Lodderomyces;Pachysolen; Pichia; Saccharomyces; Saturnispora; Tetrapisispora;Torulaspora; Williopsis; and Zygosaccharomyces. Other types of yeastpotentially useful in the invention include Yarrowia; Rhodosporidium;Candida; Hansenula; Filobasium; Sporidiobolus; Bullera; Leucosporidiumand Filobasidella.

In a preferred embodiment of the invention, the mating competent yeastis a member of the genus Pichia. In a further preferred embodiment ofthe invention, the mating competent yeast of the genus Pichia is one ofthe following species: Pichia pastoris, Pichia methanolica, andHansenula polymorphs (Pichia angusta). In a particularly preferredembodiment of the invention, the mating competent yeast of the genusPichia is the species Pichia pastoris.

Haploid Yeast Cell:

A cell having a single copy of each gene of its normal genomic(chromosomal) complement.

Polyploid Yeast Cell:

A cell having more than one copy of its normal genomic (chromosomal)complement.

Diploid Yeast Cell:

A cell having two copies (alleles) of essentially every gene of itsnormal genomic complement, typically formed by the process of fusion(mating) of two haploid cells.

Tetraploid Yeast Cell:

A cell having four copies (alleles) of essentially every gene of itsnormal genomic complement, typically formed by the process of fusion(mating) of two haploid cells. Tetraploids may carry two, three, four ormore different expression cassettes. Such tetraploids might be obtainedin S. cerevisiae by selective mating homozygotic heterothallic a/a andalpha/alpha diploids and in Pichia by sequential mating of haploids toobtain auxotrophic diploids. For example, a [met his] haploid can bemated with [ade his] haploid to obtain diploid [his]; and a [met arg]haploid can be mated with [ade arg] haploid to obtain diploid [arg];then the diploid [his]×diploid [arg] to obtain a tetraploid prototroph.It will be understood by those of skill in the art that reference to thebenefits and uses of diploid cells may also apply to tetraploid cells.

Yeast Mating:

The process by which two haploid yeast cells naturally fuse to form onediploid yeast cell.

Meiosis:

The process by which a diploid yeast cell undergoes reductive divisionto form four haploid spore products. Each spore may then germinate andform a haploid vegetatively growing cell line.

Selectable Marker:

A selectable marker is a gene or gene fragment that confers a growthphenotype (physical growth characteristic) on a cell receiving that geneas, for example through a transformation event. The selectable markerallows that cell to survive and grow in a selective growth medium underconditions in which cells that do not receive that selectable markergene cannot grow. Selectable marker genes generally fall into severaltypes, including positive selectable marker genes such as a gene thatconfers on a cell resistance to an antibiotic or other drug, temperaturewhen two temperature sensitive (“ts”) mutants are crossed or a is mutantis transformed; negative selectable marker genes such as a biosyntheticgene that confers on a cell the ability to grow in a medium without aspecific nutrient needed by all cells that do not have that biosyntheticgene, or a mutagenized biosynthetic gene that confers on a cellinability to grow by cells that do not have the wild type gene; and thelike. Suitable markers include but are not limited to: ZEO; G418; LYS3;MET1; MET3a; ADE1; ADE3; URA3; and the like.

Expression Vector:

These DNA vectors contain elements that facilitate manipulation for theexpression of a foreign protein within the target host cell.Conveniently, manipulation of sequences and production of DNA fortransformation is first performed in a bacterial host, e.g. E. coli, andusually vectors will include sequences to facilitate such manipulations,including a bacterial origin of replication and appropriate bacterialselection marker. Selection markers encode proteins necessary for thesurvival or growth of transformed host cells grown in a selectiveculture medium. Host cells not transformed with the vector containingthe selection gene will not survive in the culture medium. Typicalselection genes encode proteins that (a) confer resistance toantibiotics or other toxins, (b) complement auxotrophic deficiencies, or(c) supply critical nutrients not available from complex media.Exemplary vectors and methods for transformation of yeast are described,for example, in Burke, D., Dawson, D., & Stearns, T. (2000). Methods inyeast genetics: a Cold Spring Harbor Laboratory course manual.Plainview, N.Y.: Cold Spring Harbor Laboratory Press.

Expression vectors for use in the methods of the invention will furtherinclude yeast specific sequences, including a selectable auxotrophic ordrug marker for identifying transformed yeast strains. A drug marker mayfurther be used to amplify copy number of the vector in a yeast hostcell.

The polypeptide coding sequence of interest is operably linked totranscriptional and translational regulatory sequences that provide forexpression of the polypeptide in yeast cells. These vector componentsmay include, but are not limited to, one or more of the following: anenhancer element, a promoter, and a transcription termination sequence.Sequences for the secretion of the polypeptide may also be included,e.g. a signal sequence, and the like. A yeast origin of replication isoptional, as expression vectors are often integrated into the yeastgenome. In one embodiment of the invention, the polypeptide of interestis operably linked, or fused, to sequences providing for optimizedsecretion of the polypeptide from yeast diploid cells.

Nucleic acids are “operably linked” when placed into a functionalrelationship with another nucleic acid sequence. For example, DNA for asignal sequence is operably linked to DNA for a polypeptide if it isexpressed as a preprotein that participates in the secretion of thepolypeptide; a promoter or enhancer is operably linked to a codingsequence if it affects the transcription of the sequence. Generally,“operably linked” means that the DNA sequences being linked arecontiguous, and, in the case of a secretory leader, contiguous and inreading frame. However, enhancers do not have to be contiguous. Linkingis accomplished by ligation at convenient restriction sites oralternatively via a PCR/recombination method familiar to those skilledin the art (Gateway® Technology; Invitrogen, Carlsbad Calif.). If suchsites do not exist, the synthetic oligonucleotide adapters or linkersare used in accordance with conventional practice.

Promoters are untranslated sequences located upstream (5′) to the startcodon of a structural gene (generally within about 100 to 1000 bp) thatcontrol the transcription and translation of particular nucleic acidsequences to which they are operably linked. Such promoters fall intoseveral classes: inducible, constitutive, and repressible promoters(that increase levels of transcription in response to absence of arepressor). Inducible promoters may initiate increased levels oftranscription from DNA under their control in response to some change inculture conditions, e.g., the presence or absence of a nutrient or achange in temperature.

The promoter fragment may also serve as the site for homologousrecombination and integration of the expression vector into the samesite in the host genome; alternatively a selectable marker is used asthe site for homologous recombination.

Examples of suitable promoters useful in Pichia include the AOX1 andpromoter (Cregg et al. (1989) Mol. Cell. Biol. 9:1316-1323); ICL1promoter (Menendez et al. (2003) Yeast 20(13): 1097-108);glyceraldehyde-3-phosphate dehydrogenase promoter (GAP) (Waterham et al.(1997) Gene 186(1):37-44); and FLD1 promoter (Shen et al. (1998) Gene216(1):93-102). The GAP promoter is a strong constitutive promoter andthe AOX and FLD1 promoters are inducible.

Other yeast promoters include ADH1, alcohol dehydrogenase II, GAL4,PHO3, PHO5, Pyk, and chimeric promoters derived therefrom. Additionally,non-yeast promoters may be used in the invention such as mammalian,insect, plant, reptile, amphibian, bacterial, fungal, viral, and avianpromoters. Most typically the promoter will comprise a mammalianpromoter (potentially endogenous to the expressed genes) or willcomprise a yeast or viral promoter that provides for efficienttranscription in yeast systems.

The polypeptides of interest may be produced recombinantly not onlydirectly, but also as a fusion polypeptide with a heterologouspolypeptide, e.g. a signal sequence or other polypeptide having aspecific cleavage site at the N-terminus of the mature protein orpolypeptide. In general, the signal sequence may be a component of thevector, or it may be a part of the polypeptide coding sequence that isinserted into the vector. The heterologous signal sequence selectedpreferably is one that is recognized and processed through one of thestandard pathways available within the host cell. The S. cerevisiaealpha factor pre-pro signal has proven effective in the secretion of avariety of recombinant proteins from P. pastoris. Other yeast signalsequences include the alpha mating factor signal sequence, the invertasesignal sequence, and signal sequences derived from other secreted yeastpolypeptides. Additionally, these signal peptide sequences may beengineered to provide for enhanced secretion in diploid yeast expressionsystems. Other secretion signals of interest also include mammaliansignal sequences, which may be heterologous to the protein beingsecreted, or may be a native sequence for the protein being secreted.Signal sequences include pre-peptide sequences, and in some instancesmay include propeptide sequences. Many such signal sequences are knownin the art, including the signal sequences found on immunoglobulinchains, e.g., K28 preprotoxin sequence, PHA-E, FACE, human MCP-1, humanserum albumin signal sequences, human Ig heavy chain, human Ig lightchain, and the like. For example, see Hashimoto et. al. Protein Eng11(2) 75 (1998); and Kobayashi et. al. Therapeutic Apheresis 2(4) 257(1998).

Transcription may be increased by inserting a transcriptional activatorsequence into the vector. These activators are cis-acting elements ofDNA, usually about from 10 to 300 bp, which act on a promoter toincrease its transcription. Transcriptional enhancers are relativelyorientation and position independent, having been found 5′ and 3′ to thetranscription unit, within an intron, as well as within the codingsequence itself. The enhancer may be spliced into the expression vectorat a position 5′ or 3′ to the coding sequence, but is preferably locatedat a site 5′ from the promoter.

Expression vectors used in eukaryotic host cells may also containsequences necessary for the termination of transcription and forstabilizing the mRNA. Such sequences are commonly available from 3′ tothe translation termination codon, in untranslated regions of eukaryoticor viral DNAs or cDNAs. These regions contain nucleotide segmentstranscribed as polyadenylated fragments in the untranslated portion ofthe mRNA.

Construction of suitable vectors containing one or more of theabove-listed components employs standard ligation techniques orPCR/recombination methods. Isolated plasmids or DNA fragments arecleaved, tailored, and re-ligated in the form desired to generate theplasmids required or via recombination methods. For analysis to confirmcorrect sequences in plasmids constructed, the ligation mixtures areused to transform host cells, and successful transformants selected byantibiotic resistance (e.g. ampicillin or Zeocin) where appropriate.Plasmids from the transformants are prepared, analyzed by restrictionendonuclease digestion and/or sequenced.

As an alternative to restriction and ligation of fragments,recombination methods based on att sites and recombination enzymes maybe used to insert DNA sequences into a vector. Such methods aredescribed, for example, by Landy (1989) Ann. Rev. Biochem. 58:913-949;and are known to those of skill in the art. Such methods utilizeintermolecular DNA recombination that is mediated by a mixture of lambdaand E. coli-encoded recombination proteins. Recombination occurs betweenspecific attachment (att) sites on the interacting DNA molecules. For adescription of att sites see Weisberg and Landy (1983) Site-SpecificRecombination in Phage Lambda, in Lambda II, Weisberg, ed. (Cold SpringHarbor, N.Y.: Cold Spring Harbor Press), pp. 211-250. The DNA segmentsflanking the recombination sites are switched, such that afterrecombination, the att sites are hybrid sequences comprised of sequencesdonated by each parental vector. The recombination can occur betweenDNAs of any topology.

Att sites may be introduced into a sequence of interest by ligating thesequence of interest into an appropriate vector; generating a PCRproduct containing att B sites through the use of specific primers;generating a cDNA library cloned into an appropriate vector containingatt sites; and the like.

Folding, as used herein, refers to the three-dimensional structure ofpolypeptides and proteins, where interactions between amino acidresidues act to stabilize the structure. While non-covalent interactionsare important in determining structure, usually the proteins of interestwill have intra- and/or intermolecular covalent disulfide bonds formedby two cysteine residues. For naturally occurring proteins andpolypeptides or derivatives and variants thereof, the proper folding istypically the arrangement that results in optimal biological activity,and can conveniently be monitored by assays for activity, e.g. ligandbinding, enzymatic activity, etc.

In some instances, for example where the desired product is of syntheticorigin, assays based on biological activity will be less meaningful. Theproper folding of such molecules may be determined on the basis ofphysical properties, energetic considerations, modeling studies, and thelike.

The expression host may be further modified by the introduction ofsequences encoding one or more enzymes that enhance folding anddisulfide bond formation, i.e. foldases, chaperonins, etc. Suchsequences may be constitutively or inducibly expressed in the yeast hostcell, using vectors, markers, etc. as known in the art. Preferably thesequences, including transcriptional regulatory elements sufficient forthe desired pattern of expression, are stably integrated in the yeastgenome through a targeted methodology.

For example, the eukaryotic PDI is not only an efficient catalyst ofprotein cysteine oxidation and disulfide bond isomerization, but alsoexhibits chaperone activity. Co-expression of PDI can facilitate theproduction of active proteins having multiple disulfide bonds. Also ofinterest is the expression of BIP (immunoglobulin heavy chain bindingprotein); cyclophilin; and the like. In one embodiment of the invention,each of the haploid parental strains expresses a distinct foldingenzyme, e.g. one strain may express BIP, and the other strain mayexpress PDI or combinations thereof.

The terms “desired protein” or “desired antibody” are usedinterchangeably and refer generally to a parent antibody or fragmentspecific to a target, i.e., PCSK9 or a chimeric or humanized antibody ora binding portion thereof derived therefrom or one containing the sameCDRs or epitopic specificity as any of the anti-PCSK9 antibodies orfragments described herein. The term “antibody” is intended to includeany polypeptide chain-containing molecular structure with a specificshape that fits to and recognizes an epitope, where one or morenon-covalent binding interactions stabilize the complex between themolecular structure and the epitope. The archetypal antibody molecule isthe immunoglobulin, and all types of immunoglobulins, IgG, IgM, IgA,IgE, IgD, etc., from all sources, e.g. human, rodent, rabbit, cow,sheep, pig, dog, other mammals, chicken, other avians, etc., areconsidered to be “antibodies.” A preferred source for producingantibodies useful as starting material according to the invention israbbits. Numerous antibody coding sequences have been described; andothers may be raised by methods well-known in the art. Examples thereofinclude chimeric antibodies, human antibodies and other non-humanmammalian antibodies, humanized antibodies, single chain antibodies(such as scFvs), camelbodies, nanobodies, IgNAR (single-chain antibodiesderived from sharks), small-modular immunopharmaceuticals (SMIPs), andantibody fragments such as Fabs, Fab′, F(ab′)₂, monovalent antibodyfragments such as MetMab like molecules, IgNars and the like. SeeStreltsov V A, et al., Structure of a shark IgNAR antibody variabledomain and modeling of an early-developmental isotype, Protein Sci. 2005November; 14(11):2901-9. Epub 2005 Sep. 30; Greenberg A S, et al., A newantigen receptor gene family that undergoes rearrangement and extensivesomatic diversification in sharks, Nature. 1995 Mar. 9;374(6518):168-73; Nuttall S D, et al., Isolation of the new antigenreceptor from wobbegong sharks, and use as a scaffold for the display ofprotein loop libraries, Mol. Immunol. 2001 August; 38(4):313-26;Hamers-Casterman C, et al., Naturally occurring antibodies devoid oflight chains, Nature. 1993 Jun. 3; 363(6428):446-8; Gill D S, et al.,Biopharmaceutical drug discovery using novel protein scaffolds, CurrOpin Biotechnol. 2006 December; 17(6):653-8. Epub 2006 Oct. 19.

The present invention includes in particular includes monovalentantibody molecules that bind PCSK9, which are analogous to MetMabmolecules. MetMab is a monovalent antibody specific to Met. (Met is aprotein encoded by the nucleotide sequence set forth in Park et al.,Proc. Natl. Acad. Sci. 84, 7479—(1987), or fragments thereof, as well asrelated polypeptides, which include, but are not limited to, allelicvariants, splice variants, derivative variants, substitution variants,deletion variants, and/or insertion variants, fusion polypeptides, andinterspecies homologs). The MetMab antibody, is a monovalent antibodyknown by different names including OA-5d5 (Genentech) and is also calledOne Armed 5d5, 5d5, MetMab, PRO143966, among others). Antibody OA-5d5,including its structure and properties, and methods for making and usingit, are described in U.S. Publication No. 2007/0092520. In oneembodiment, an anti-PCSK9 antibody according to the invention maycomprise a single Fab region linked to an Fc region. In such embodiment,an antibody of the invention may comprise light and heavy chain variabledomains as described herein. In such an embodiment, the antibody ismonovalent and may comprise an intact Fc region. In another suchembodiment, the Fc region may comprise at least one protuberance (knob)and at least one cavity (hole), wherein the presence of the protuberanceand cavity enhances formation of a complex between an Fc polypeptidecomprising the protuberance and an Fc polypeptide comprising the cavity,for example as described in WO 2005/063816. In one embodiment, the Fcregion of an antibody of the invention may comprise a first and a secondFc polypeptide, wherein the first and second polypeptide each comprisesone or more mutations with respect to wild type human Fc. In oneembodiment, a cavity mutation is T366S, L368A and/or Y407V. In anotherembodiment, a protuberance mutation is T366W. In a specific embodiment,a monovalent antibody according to the subject invention may comprise aone-armed antibody synthesized as described in WO2005/063816. In suchembodiment, the one-armed antibody may comprise Fc mutationsconstituting “knobs” and “holes” as described in WO2005/063816. Forexample, a hole mutation can be one or more of T366A, L368A and/or Y407Vin an Fc polypeptide, and a cavity mutation can be T366W. The inventionis also directed to an anti-human PCSK9 monovalent agent that binds withthe same PCSK9 epitope and/or competes with an anti-PCSK9 antibody forbinding to PCSK9 as an antibody or antibody fragment disclosed herein.

For example, antibodies or antigen binding fragments may be produced bygenetic engineering. In this technique, as with other methods,antibody-producing cells are sensitized to the desired antigen orimmunogen. The messenger RNA isolated from antibody producing cells isused as a template to make cDNA using PCR amplification. A library ofvectors, each containing one heavy chain gene and one light chain generetaining the initial antigen specificity, is produced by insertion ofappropriate sections of the amplified immunoglobulin cDNA into theexpression vectors. A combinatorial library is constructed by combiningthe heavy chain gene library with the light chain gene library. Thisresults in a library of clones which co-express a heavy and light chain(resembling the Fab fragment or antigen binding fragment of an antibodymolecule). The vectors that carry these genes are co-transfected into ahost cell. When antibody gene synthesis is induced in the transfectedhost, the heavy and light chain proteins self-assemble to produce activeantibodies that can be detected by screening with the antigen orimmunogen.

Antibody coding sequences of interest include those encoded by nativesequences, as well as nucleic acids that, by virtue of the degeneracy ofthe genetic code, are not identical in sequence to the disclosed nucleicacids, and variants thereof. Variant polypeptides can include amino acid(aa) substitutions, additions or deletions. The amino acid substitutionscan be conservative amino acid substitutions or substitutions toeliminate non-essential amino acids, such as to alter a glycosylationsite, or to minimize misfolding by substitution or deletion of one ormore cysteine residues that are not necessary for function. Variants canbe designed so as to retain or have enhanced biological activity of aparticular region of the protein (e.g., a functional domain, catalyticamino acid residues, etc). Variants also include fragments of thepolypeptides disclosed herein, particularly biologically activefragments and/or fragments corresponding to functional domains.Techniques for in vitro mutagenesis of cloned genes are known. Alsoincluded in the subject invention are polypeptides that have beenmodified using ordinary molecular biological techniques so as to improvetheir resistance to proteolytic degradation or to optimize solubilityproperties or to render them more suitable as a therapeutic agent.

Chimeric antibodies may be made by recombinant means by combining thevariable light and heavy chain regions (V_(L) and V_(H)), obtained fromantibody producing cells of one species with the constant light andheavy chain regions from another. Typically chimeric antibodies utilizerodent or rabbit variable regions and human constant regions, in orderto produce an antibody with predominantly human domains. The productionof such chimeric antibodies is well known in the art, and may beachieved by standard means (as described, e.g., in U.S. Pat. No.5,624,659, incorporated herein by reference in its entirety). It isfurther contemplated that the human constant regions of chimericantibodies of the invention may be selected from IgG1, IgG2, IgG3, andIgG4 constant regions.

Humanized antibodies are engineered to contain even more human-likeimmunoglobulin domains, and incorporate only thecomplementarity-determining regions of the animal-derived antibody. Thisis accomplished by carefully examining the sequence of thehyper-variable loops of the variable regions of the monoclonal antibody,and fitting them to the structure of the human antibody chains. Althoughfacially complex, the process is straightforward in practice. See, e.g.,U.S. Pat. No. 6,187,287, incorporated fully herein by reference.

In addition to entire immunoglobulins (or their recombinantcounterparts), immunoglobulin fragments comprising the epitope bindingsite (e.g., Fab′, F(ab′)₂, Fab, or other fragments) may be synthesized.“Fragment” or minimal immunoglobulins may be designed utilizingrecombinant immunoglobulin techniques. For instance “Fv” immunoglobulinsfor use in the present invention may be produced by synthesizing a fusedvariable light chain region and a variable heavy chain region.Combinations of antibodies are also of interest, e.g. diabodies, whichcomprise two distinct Fv specificities. In another embodiment of theinvention, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, and IgNAR are encompassed by immunoglobulin fragments.

Immunoglobulins and fragments thereof may be modifiedpost-translationally, e.g. to add effector moieties such as chemicallinkers, detectable moieties, such as fluorescent dyes, enzymes, toxins,substrates, bioluminescent materials, radioactive materials,chemiluminescent moieties and the like, or specific binding moieties,such as streptavidin, avidin, or biotin, and the like may be utilized inthe methods and compositions of the present invention. Examples ofadditional effector molecules are provided infra.

A polynucleotide sequence “corresponds” to a polypeptide sequence iftranslation of the polynucleotide sequence in accordance with thegenetic code yields the polypeptide sequence (i.e., the polynucleotidesequence “encodes” the polypeptide sequence), one polynucleotidesequence “corresponds” to another polynucleotide sequence if the twosequences encode the same polypeptide sequence.

A “heterologous” region or domain of a DNA construct is an identifiablesegment of DNA within a larger DNA molecule that is not found inassociation with the larger molecule in nature. Thus, when theheterologous region encodes a mammalian gene, the gene will usually beflanked by DNA that does not flank the mammalian genomic DNA in thegenome of the source organism. Another example of a heterologous regionis a construct where the coding sequence itself is not found in nature(e.g., a cDNA where the genomic coding sequence contains introns, orsynthetic sequences having codons different than the native gene).Allelic variations or naturally-occurring mutational events do not giverise to a heterologous region of DNA as defined herein.

A “coding sequence” is an in-frame sequence of codons that (in view ofthe genetic code) correspond to or encode a protein or peptide sequence.Two coding sequences correspond to each other if the sequences or theircomplementary sequences encode the same amino acid sequences. A codingsequence in association with appropriate regulatory sequences may betranscribed and translated into a polypeptide. A polyadenylation signaland transcription termination sequence will usually be located 3′ to thecoding sequence. A “promoter sequence” is a DNA regulatory regioncapable of binding RNA polymerase in a cell and initiating transcriptionof a downstream (3′ direction) coding sequence. Promoter sequencestypically contain additional sites for binding of regulatory molecules(e.g., transcription factors) which affect the transcription of thecoding sequence. A coding sequence is “under the control” of thepromoter sequence or “operatively linked” to the promoter when RNApolymerase binds the promoter sequence in a cell and transcribes thecoding sequence into mRNA, which is then in turn translated into theprotein encoded by the coding sequence.

Vectors are used to introduce a foreign substance, such as DNA, RNA orprotein, into an organism or host cell. Typical vectors includerecombinant viruses (for polynucleotides) and liposomes (forpolypeptides). A “DNA vector” is a replicon, such as plasmid, phage orcosmid, to which another polynucleotide segment may be attached so as tobring about the replication of the attached segment. An “expressionvector” is a DNA vector which contains regulatory sequences which willdirect polypeptide synthesis by an appropriate host cell. This usuallymeans a promoter to bind RNA polymerase and initiate transcription ofmRNA, as well as ribosome binding sites and initiation signals to directtranslation of the mRNA into a polypeptide(s). Incorporation of apolynucleotide sequence into an expression vector at the proper site andin correct reading frame, followed by transformation of an appropriatehost cell by the vector, enables the production of a polypeptide encodedby said polynucleotide sequence.

“Amplification” of polynucleotide sequences is the in vitro productionof multiple copies of a particular nucleic acid sequence. The amplifiedsequence is usually in the form of DNA. A variety of techniques forcarrying out such amplification are described in a review article by VanBrunt (1990, Bio/Technol., 8(4):291-294). Polymerase chain reaction orPCR is a prototype of nucleic acid amplification, and use of PCR hereinshould be considered exemplary of other suitable amplificationtechniques.

The general structure of antibodies in vertebrates now is wellunderstood (Edelman, G. M., Ann. N.Y. Acad. Sci., 190: 5 (1971)).Antibodies consist of two identical light polypeptide chains ofmolecular weight approximately 23,000 Daltons (the “light chain”), andtwo identical heavy chains of molecular weight 53,000-70,000 (the “heavychain”). The four chains are joined by disulfide bonds in a “Y”configuration wherein the light chains bracket the heavy chains startingat the mouth of the “Y” configuration. The “branch” portion of the “Y”configuration is designated the F_(ab) region; the stem portion of the“Y” configuration is designated the F_(c) region. The amino acidsequence orientation runs from the N-terminal end at the top of the “Y”configuration to the C-terminal end at the bottom of each chain. TheN-terminal end possesses the variable region having specificity for theantigen that elicited it, and is approximately 100 amino acids inlength, there being slight variations between light and heavy chain andfrom antibody to antibody.

The variable region is linked in each chain to a constant region thatextends the remaining length of the chain and that within a particularclass of antibody does not vary with the specificity of the antibody(i.e., the antigen eliciting it). There are five known major classes ofconstant regions that determine the class of the immunoglobulin molecule(IgG, IgM, IgA, IgD, and IgE corresponding to γ, μ, α, δ, and ε (gamma,mu, alpha, delta, or epsilon) heavy chain constant regions). Theconstant region or class determines subsequent effector function of theantibody, including activation of complement (Kabat, E. A., StructuralConcepts in Immunology and Immunochemistry, 2nd Ed., p. 413-436, Holt,Rinehart, Winston (1976)), and other cellular responses (Andrews, D. W.,et al., Clinical Immunobiology, pp 1-18, W. B. Sanders (1980); Kohl, S.,et al., Immunology, 48: 187 (1983)); while the variable regiondetermines the antigen with which it will react. Light chains areclassified as either κ (kappa) or λ (lambda). Each heavy chain class canbe prepared with either kappa or lambda light chain. The light and heavychains are covalently bonded to each other, and the “tail” portions ofthe two heavy chains are bonded to each other by covalent disulfidelinkages when the immunoglobulins are generated either by hybridomas orby B cells.

The expression “variable region” or “VR” refers to the domains withineach pair of light and heavy chains in an antibody that are involveddirectly in binding the antibody to the antigen. Each heavy chain has atone end a variable domain (V_(H)) followed by a number of constantdomains. Each light chain has a variable domain (V_(L)) at one end and aconstant domain at its other end; the constant domain of the light chainis aligned with the first constant domain of the heavy chain, and thelight chain variable domain is aligned with the variable domain of theheavy chain.

The expressions “complementarity determining region,” “hypervariableregion,” or “CDR” refer to one or more of the hyper-variable orcomplementarity determining regions (CDRs) found in the variable regionsof light or heavy chains of an antibody (See Kabat, E. A. et al.,Sequences of Proteins of Immunological Interest, National Institutes ofHealth, Bethesda, Md., (1987)). These expressions include thehypervariable regions as defined by Kabat et al. (“Sequences of Proteinsof Immunological Interest,” Kabat E., et al., US Dept. of Health andHuman Services, 1983) or the hypervariable loops in 3-dimensionalstructures of antibodies (Chothia and Lesk, J. Mol. Biol. 196 901-917(1987)). The CDRs in each chain are held in close proximity by frameworkregions and, with the CDRs from the other chain, contribute to theformation of the antigen binding site. Within the CDRs there are selectamino acids that have been described as the selectivity determiningregions (SDRs) which represent the critical contact residues used by theCDR in the antibody-antigen interaction (Kashmiri, S., Methods, 36:25-34(2005)).

An “epitope” or “binding site” is an area or region on an antigen towhich an antigen-binding peptide (such as an antibody) specificallybinds. A protein epitope may comprise amino acid residues directlyinvolved in the binding (also called immunodominant component of theepitope) and other amino acid residues, which are not directly involvedin the binding, such as amino acid residues which are effectivelyblocked by the specifically antigen binding peptide (in other words, theamino acid residue is within the “footprint” of the specifically antigenbinding peptide). The term epitope herein includes both types of aminoacid binding sites in any particular region of PCSK9 that specificallybinds to an anti-PCSK9 antibody. PCSK9 may comprise a number ofdifferent epitopes, which may include, without limitation, (1) linearpeptide antigenic determinants, (2) conformational antigenicdeterminants which consist of one or more non-contiguous amino acidslocated near each other in a mature PCSK9 conformation; and (3)post-translational antigenic determinants which consist, either in wholeor part, of molecular structures covalently attached to a PCSK9 proteinsuch as carbohydrate groups.

The phrase that a first antibody binds “substantially” or “at leastpartially” the same epitope as a second antibody means that the epitopebinding site for the first antibody comprises at least 10%, 20%, 30%,40%, 50%, 60%, 70%, 80%, 90%, or more of the amino acid residues on theantigen that constitutes the epitope binding site of the secondantibody. Also, that a first antibody binds substantially or partiallythe same or overlapping epitope as a second antibody means that thefirst and second antibodies compete in binding to the antigen, asdescribed above. Thus, the term “binds to substantially the same epitopeor determinant as” a monoclonal antibody means that an antibody“competes” with the antibody.

The phrase “binds to the same or overlapping epitope or determinant as”an antibody of interest means that an antibody “competes” with saidantibody of interest for at least one, or all residues on PCSK9 to whichsaid antibody of interest specifically binds. The identification of oneor more antibodies that bind(s) to substantially or essentially the sameepitope as the monoclonal antibodies described herein can be readilydetermined using any one of variety of immunological screening assays inwhich antibody competition can be assessed. A number of such assays areroutinely practiced and well known in the art (see, e.g., U.S. Pat. No.5,660,827, issued Aug. 26, 1997, which is specifically incorporatedherein by reference). It will be understood that actually determiningthe epitope to which an antibody described herein binds is not in anyway required to identify an antibody that binds to the same orsubstantially the same or overlapping epitope as the monoclonal antibodydescribed herein.

For example, where the test antibodies to be examined are obtained fromdifferent source animals, or are even of a different Ig isotype, asimple competition assay may be employed in which the control antibodyis mixed with the test antibody and then applied to a sample containingPCSK9. Protocols based upon ELISAs, radioimmunoassays, Western blotting,and the use of BIACORE analysis are suitable for use in such simplecompetition studies.

In certain embodiments, one would pre-mix the control anti-PCSK9antibody with varying amounts of the test antibody (e.g., in ratios ofabout 1:1, 1:2, 1:10 or about 1:100) for a period of time prior toapplying to the PCSK9 antigen sample. In other embodiments, the controland varying amounts of test antibody can simply be added separately andadmixed during exposure to the PCSK9 antigen sample. As long as one candistinguish bound from free antibodies (e.g., by using separation orwashing techniques to eliminate unbound antibodies) and control antibodyfrom the test antibody (e.g., by using species specific or isotypespecific secondary antibodies or by specifically labeling the controlantibody with a detectable label) one will be able to determine if thetest antibody reduces the binding of the control antibody to the PCSK9antigens, indicating that the test antibody recognizes substantially thesame epitope as the control ant-PCSK9 antibody. The binding of the(labeled) control antibody in the presence of a completely irrelevantantibody (that does not bind PCSK9) can serve as the control high value.The control low value can be obtained by incubating the labeled controlantibody with the same but unlabeled control antibody, where competitionwould occur and reduce binding of the labeled antibody. In a test assay,a significant reduction in labeled antibody reactivity in the presenceof a test antibody is indicative of a test antibody that recognizessubstantially the same epitope, i.e., one that competes with the labeledcontrol antibody. For example, any test antibody that reduces thebinding of the control antibody to PCSK9 s by at least about 50%, suchas at least about 60%, or more preferably at least about 70% (e.g.,about 65-100%), at any ratio of: test antibody between about 1:1 or 1:10and about 1:100 is considered to be an antibody that binds tosubstantially the same or overlapping epitope or determinant as thecontrol antibody.

Preferably, such test antibody will reduce the binding of the controlantibody to PCSK9 antigen preferably at least about 50%, at least about60%, at least about 80% or at least about 90% (e.g., about 95%) of thebinding of 1 the control antibody observed in the absence of the testantibody.

Competition can also or alternatively be assessed by, for example, aflow cytometry test. In such a test, cells bearing PCSK9 can beincubated first with a control antibody that binds PCSK9, and then withthe test antibody labeled with a fluorochrome or biotin. The antibody issaid to compete with control antibody if the binding obtained uponpreincubation with saturating amount of control antibody is about 80%,preferably about 50%, about 40% or less (e.g., about 30%) of the binding(as measured by mean of fluorescence) obtained by the test antibodywithout preincubation with control antibody. Alternatively, an antibodyis said to compete with the control antibody if the binding obtainedwith a labeled control antibody (by a fluorochrome or biotin) on cellspreincubated with saturating amount of test antibody is about 80%,preferably about 50%, about 40%, or less (e.g., about 30%) of thebinding obtained without preincubation with the test antibody.

A simple competition assay in which a test antibody is pre-adsorbed andapplied at saturating concentration to a surface onto which PCSK9 isimmobilized also may be advantageously employed. The surface in thesimple competition assay is preferably a BIACORE chip (or other mediasuitable for surface plasmon resonance analysis). The binding of acontrol antibody that binds PCSK9 to the PCSK9-coated surface ismeasured. This binding to the PCSK9-containing surface of the controlantibody alone is compared with the binding of the control antibody inthe presence of a test antibody. A significant reduction in binding tothe PCSK9-containing surface by the control antibody in the presence ofa test antibody indicates that the test antibody recognizessubstantially the same epitope as the control antibody such that thetest antibody “competes” with the control antibody. Any test antibodythat reduces the binding of control antibody by at least about 20% ormore, at least about 40%, at least about 50%, at least about 70%, ormore, can be considered to be an antibody that binds to substantiallythe same epitope or determinant as the control antibody. Preferably,such test antibody will reduce the binding of the control antibody toPCSK9 by at least about 50% (e.g., at least about 60%, at least about70%, or more). It will be appreciated that the order of control and testantibodies can be reversed; i.e. the control antibody can be first boundto the surface and then the test antibody is brought into contact withthe surface thereafter in a competition assay. Preferably, the antibodyhaving higher affinity for PCSK9 antigen is bound to thePCSK9-containing surface first, as it will be expected that the decreasein binding seen for the second antibody (assuming the antibodies arecompeting) will be of greater magnitude. Further examples of such assaysare provided in e.g., Saunal and Regenmortel, (1995) J. Immunol. Methods183: 33-41, the disclosure of which is incorporated herein by reference.

In addition, whether an antibody binds the same or overlappingepitope(s) on PCSK9 as another antibody or the epitope bound by a testantibody may in particular be determined using a western-blot basedassay. In this assay a library of peptides corresponding to the antigenbound by the antibody, herein PCSK9 is made, which correspond tooverlapping portions of the protein, typically 10-25, 10-20 or 10-15amino acids long. These different overlapping amino acid peptidesencompassing the PCSK9 sequence are synthesized and covalently bound toa PepSpots nitrocellulose membrane (JPT Peptide technologies, Berlin,Germany). Blots are then prepared and probed according to themanufacturer's recommendations.

Essentially, the immunoblot assay then detects by fluorimetric meanswhat peptides in the library bind to the test antibody and thereby canidentify what residues on the antigen, i.e., PCSK9, interact with thetest antibody. (See an embodiment of this technique in U.S. Pat. No.7,935,340, incorporated by reference herein).

The expressions “framework region” or “FR” refer to one or more of theframework regions within the variable regions of the light and heavychains of an antibody (See Kabat, E. A. et al., Sequences of Proteins ofImmunological Interest, National Institutes of Health, Bethesda, Md.,(1987)). These expressions include those amino acid sequence regionsinterposed between the CDRs within the variable regions of the light andheavy chains of an antibody.

Anti-PCSK9 Antibodies and Binding Fragments Thereof Having BindingActivity for PCSK9

As mentioned in the Background of the invention, Proprotein convertasesubtilisin kexin type 9 (PCSK9) is a serine protease involved inregulating the levels of the low density lipoprotein receptor (LDLR)protein (Horton et al., 32(2) Trends Biochem. Sci. 71-77 (2007); Seidahand Prat, 85(7) J. Mol. Med. 685-96 2007). PCSK9 is aprohormone-proprotein convertase in the subtilisin (S8) family of serineproteases Seidah et al., 100(3) Proc. Nat'l Acad. Sci. 928-33 (2003).Exemplary human PCSK9 amino acid sequences are presented as SEQ ID NO:961 and SEQ ID NO: 962. An exemplary human PCSK9 coding sequence ispresented as SEQ ID NO: 963. As described herein, PCSK9 proteins canalso include fragments of the full length PCSK9 protein. The structureof the PCSK9 protein has recently been solved by two groups (Cunninghamet al., Nature Structural & Molecular Biology, 2007, and Piper et al.,Structure, 15:1-8, 2007), the entireties of both of which are hereinincorporated by reference. PCSK9 includes a signal sequence, aN-terminal prodomain, a subtilisin-like catalytic domain, and aC-terminal domain.

The present invention provides novel antibodies or antibody fragmentsthat bind PCSK9, including human PCSK9. In preferred embodiments, theantibody or antibody fragment according to the invention comprises oneor more complementarity determining regions (CDRs), of the anti-PCSK9antibodies and antibody fragments described herein.

In some embodiments, an anti-PCSK9 antibody or antibody fragmentaccording to the invention will interfere with, block, reduce ormodulate the interaction between PCSK9 and LDLR. In some instances ananti-PCSK9 antibody or antibody fragment according to the invention isdenoted as “neutralizing”, e.g., if it totally prevents the interactionof PCSK9 and LDLR. In some embodiments, the antibody or antibodyfragment neutralizes PCSK9, e.g., by remaining bound to PCSK9. Forexample, in some embodiments, the antibody or antibody fragmentaccording to the invention prevents or reduces the adverse influence ofPCSK9 on LDLR without blocking the LDLR binding site on PCSK9. Thus, insome embodiments, the antibody or antibody fragment according to theinvention modulates or alters PCSK9's ability to result in thedegradation of LDLR, without having to prevent the binding interactionbetween PCSK9 and LDLR. More specifically, such an antibody or antibodyfragment according to the invention can be specifically described as a“non-competitively neutralizing” antibody or antibody fragment. In someembodiments, the neutralizing antibody or antibody fragment according tothe invention binds to PCSK9 in a location and/or manner that preventsPCSK9 from binding to LDLR. In some embodiments, the neutralizingantibody or antibody fragment according to the invention binds to PCSK9in a location and/or manner that prevents endocytosis of the PCSK9/LDLRcomplex. Such antibody or antibody fragment according to the inventioncan be specifically described as “competitively neutralizing” antibodyor antibody fragment according to the invention. All of the aboveneutralizing antibodies upon in vivo administration may result in agreater amount of free LDLR being present in a subject, which results inmore LDLR binding to LDL-C (thereby reducing the amount of LDL-C in thesubject). This in turn this results in a reduction in the amount ofserum cholesterol present in a subject.

In some embodiments, the antibody or antibody fragment according to theinvention are capable of inhibiting PCSK9-mediated activity (includingbinding). In some embodiments, the antibody or antibody fragmentaccording to the invention are human, such as fully human antibodies toPCSK9.

In some embodiments, the antibody or antibody fragment according to theinvention binds to the catalytic domain of PCSK9. In some embodiments,the antibody or antibody fragment according to the invention binds tothe mature form of PCSK9. In some embodiments the antibody or antibodyfragment according to the invention binds in the prodomain of PCSK9. Insome embodiments, the antibody or antibody fragment according to theinvention selectively binds to the mature form of PCSK9. In someembodiments, the antibody or antibody fragment according to theinvention binds to the catalytic domain in a manner such that PCSK9cannot bind or bind as efficiently to LDLR. In some embodiments, theantibody or antibody fragment according to the invention does not bindto the c-terminus of the catalytic domain. In some embodiments, theantibody or antibody fragment according to the invention does not bindto the n-terminus of the catalytic domain. In some embodiments, theantibody or antibody fragment according to the invention does not bindto the n- or c-terminus of the PCSK9 protein. In some embodiments, theantibody or antibody fragment according to the invention bind to aspecific conformational state of PCSK9 so as to prevent PCSK9 frominteracting with LDLR. In some embodiments, the antibody or antibodyfragment according to the invention binds to the V domain of PCSK9. Insome more specific embodiments, the antibody or antibody fragmentaccording to the invention binds to the V domain of PCSK9 and prevents(or reduces) PCSK9 from binding to LDLR.

As mentioned, the anti-PCSK9 antibodies or antibody fragments accordingto the invention have a variety of utilities. For example, the subjectantibodies and fragments are useful in therapeutic applications, as wellas diagnostically in binding assays, and are useful for affinitypurification of PCSK9, in particular human PCSK9 or its ligands and inscreening assays to identify other antagonists of PCSK9 activity. Someof the antibodies or antibody fragments according to the invention areuseful for inhibiting binding of PCSK9 to LDLR, or inhibitingPCSK9-mediated activities.

The antibody or antibody fragment according to the invention can be usedin a variety of therapeutic applications. For example, in someembodiments the PCSK9 antibody or antibody fragment according to theinvention are useful for treating conditions associated with PCSK9, suchas cholesterol related disorders (or “serum cholesterol relateddisorders”) including by way of example hypercholesterolemia, as furtherdescribed herein.

The subject anti-PCSK9 antibodies and antibody fragments according tothe invention can in particular be used for treating any subject whereinblocking, inhibiting or neutralizing the in vivo effect of PCSK9 orblocking or inhibiting the interaction of PCSK9 and LDLR istherapeutically desirable, wherein the subject anti-PCSK9 antibodies orantibody fragments may be used alone or in association with other activeagents or drugs.

The subject anti-PCSK9 antibodies and antibody fragments according tothe invention can also in particular be used for treating or preventingdisorders of cholesterol or lipid homeostasis and disorders which may beassociated therewith including by way of example hypercholesterolemia,hyperlipidemia, hypertriglyceridaemia, sitosterolemia, atherosclerosis,arteriosclerosis, coronary heart disease, metabolic syndrome, acutecoronary syndrome, vascular inflammation, diabetes, obesity, angina,hypertension and xanthoma by the administration of the subjectanti-PCSK9 antibodies and antibody fragments that specifically bind toPCSK9, wherein the subject antibodies and antibody fragments may be usedalone or in association with other active agents.

The subject anti-PCSK9 antibodies and antibody fragments according tothe invention can also in particular be used for preventing or treatingdiseases and disorders associated with PCSK9, e.g., diseases associatedwith increased or decreased levels of PCSK9 and/or mutations in thePCSK9 gene that affect PCSK9 protein expression, primary sequence and/orfunction by administering said antibodies or fragments thereof alone orin combination with other active agents.

The subject anti-PCSK9 antibodies and antibody fragments according tothe invention can also in particular be used for treating any subjecthaving a condition or at risk of developing a condition whereinmodulation of lipid or cholesterol levels is clinically desirable orwhere the subject has a condition that is often associated with highlipids or cholesterol.

The subject anti-PCSK9 antibodies and antibody fragments according tothe invention can also in particular be used for treating or preventingdisorders of cholesterol or lipid homeostasis and disorders andcomplications associated therewith, e.g., hypercholesterolemia,hyperlipidemia, hypertriglyceridaemia, sitosterolemia, atherosclerosis,arteriosclerosis, coronary heart disease, metabolic syndrome, acutecoronary syndrome, vascular inflammation, xanthoma, hypertension, anginaand related conditions by administration of the subject anti-PCSK9antibodies and antibody fragments alone or in association with otheractive agents.

The subject anti-PCSK9 antibodies and antibody fragments according tothe invention can also in particular be used for improving bloodcholesterol markers associated with increased risk of heart diseaseusing the subject antibodies and antibody fragments alone or inassociation with other active agents. These markers include, but are notlimited to, high total cholesterol, high LDL, high total cholesterol toHDL ratio and high LDL-C to HDL ratio.

The subject anti-PCSK9 antibodies and antibody fragments according tothe invention can also in particular be used in any of theaforementioned therapeutic indications or conditions in combination withother drugs that are typically used to treat such disorders, wherein theantibody and other drug or agent may be co-administered or separatelyadministered. Examples of drugs include that may be co-administered withthe subject anti-PCSK9 antibodies or antibody fragments or in the sametherapeutic regimen include by way of example statins, ACE inhibitors,Angiotensin II receptor blockers (ARBs), Antiarrhythmics, AntiplateletDrugs, aspirin, beta blockers, amiodarone, digoxin, aspirin,anti-clotting agents, digoxin, diuretics, heart failure drugs,vasodilators, blood thinners, other anti-cholesterol drugs such asholestyramine (Questran), gemfibrozil (Lopid, Gemcor), Omacor, andpantethine, other anti-hypertensives, antidiabetigenic drugs such asAlpha-glucosidase inhibitors, Biguanides, Dipeptidyl peptidase-4inhibitors, Insulin therapies, Meglitinides, Sulfonylurea, andThiazolidinediones, and other drugs used to treat conditions wherein thetreated individual may have high cholesterol or aberrant lipid levels orlipid metabolism.

The invention further relates to compositions containing the subjectanti-PCSK9 antibodies or antibody fragments, especially compositions aresuitable for in vivo administration, e.g., subcutaneous, intravenous,intradermal, intranasal, intrathecal, vaginal, rectal, and otherinjectable or topical administrable dosage forms.

More specifically, the invention provides compositions containing thesubject anti-PCSK9 antibodies or antibody fragments, especiallycompositions which are suitable for in vivo administration, e.g.,subcutaneous, intravenous, intradermal, intranasal, intrathecal,vaginal, rectal, oral and other injectable or topical dosage forms whichoptionally may contain another active agent such as statins, ACEinhibitors, Angiotensin II receptor blockers (ARBs), Antiarrhythmics,Antiplatelet Drugs, aspirin, beta blockers, amiodarone, digoxin,aspirin, anti-clotting agents, digoxin, diuretics, heart failure drugs,vasodilators, blood thinners, other anti-cholesterol drugs such asholestyramine (Questran), gemfibrozil (Lopid, Gemcor), Omacor, andpantethine, other anti-hypertensives, antidiabetigenic drugs such asAlpha-glucosidase inhibitors, Biguanides, Dipeptidyl peptidase-4inhibitors, Insulin therapies, Meglitinides, Sulfonylurea, andThiazolidinediones, and other drugs used to treat conditions wherein thetreated individual may have high cholesterol. The invention alsoprovides novel dosage regimens using the subject anti-PCSK9 antibodiesor antibody fragments, alone or in association with another active,especially subcutaneous, oral and intravenous dosing regimens.

Other uses for the antibodies or antibody fragments according to theinvention include, for example, diagnosis of PCSK9-associated diseasesor conditions and screening assays to determine the presence or absenceof PCSK9. Some of the antibody or antibody fragment according to theinvention described herein are useful in treating consequences,symptoms, and/or the pathology associated with PCSK9 activity.

Exemplary anti-PCSK9 antibodies and antibody fragments according to theinvention, and the specific CDRs thereof are identified in the followingsection. For the reader's convenience, each exemplified antibody orfragment, and sequences contained therein, are separately describedunder a Header that identifies the exemplified antibody by a specificnomenclature, e.g., Ab1, Ab2 and the like.

Antibody Ab1

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 1) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYWMTWVRQAPGKGLEYIGIISSSGSTYYATWAKGRFTISKTSSTTVDLEITSPTTEDTATYFCARDSAFSSGLEFNIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 2) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYWMTWVRQAPGKGLEYIGIISSSGSTYYATWAKGRFTISKTSSTTVDLEITSPTTEDTATYFCARDSAFS SGLEFNIWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab1 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 10) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 21) AYDLTQTPASVEVAVGGTVTIKCQASQSVYSNWLSWYQQKPGQPPKLLIYDASDLASGVPSRFKGSGSGTQFTLTISGVQCDDAATYYCQQGQSSSDIDNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 22) AYDLTQTPASVEVAVGGTVTIKCQASQSVYSNWLSWYQQKPGQPPKLLIYDASDLASGVPSRFKGSGSGTQFTLTISGVQCDDAATYYCQQGQSSSDID NTFGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab1 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 30) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 4; SEQ ID NO: 6; andSEQ ID NO: 8 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain sequence of SEQ IDNO: 1 or which contain the variable heavy chain sequence of SEQ ID NO:2, and/or which further contain one, two, or three of the polypeptidesequences of SEQ ID NO: 24; SEQ ID NO: 26; and SEQ ID NO: 28 whichcorrespond to the complementarity-determining regions (CDRs, orhypervariable regions) of the light chain sequence of SEQ ID NO: 21 orwhich contain the variable light chain sequence of SEQ ID NO: 22, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 3; SEQ ID NO: 5; SEQ ID NO: 7; and SEQ ID NO: 9which correspond to the framework regions (FRs or constant regions) ofthe heavy chain sequence of SEQ ID NO: 1 or the variable heavy chainsequence of SEQ ID NO: 2, and/or one, two, three, or four of thepolypeptide sequences of SEQ ID NO: 23; SEQ ID NO: 25; SEQ ID NO: 27;and SEQ ID NO: 29 which correspond to the framework regions (FRs orconstant regions) of the light chain sequence of SEQ ID NO: 21 or thevariable light chain sequence of SEQ ID NO: 22, or combinations of thesepolypeptide sequences or sequences which are at least 80%, 90%, 95%,96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 1 or SEQ ID NO: 2 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 21 orSEQ ID NO: 22 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 4; SEQ ID NO: 6; and SEQ ID NO: 8 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 1 or the variable heavy chainsequence of SEQ ID NO: 2 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 24; SEQ ID NO: 26; and SEQ ID NO: 28 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 21 or the variable light chainsequence of SEQ ID NO: 22 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 3; SEQ ID NO: 5; SEQ ID NO: 7; and SEQ ID NO: 9 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 1 or the variable heavy chainsequence of SEQ ID NO: 2 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 23; SEQ ID NO: 25; SEQ ID NO: 27; and SEQ ID NO:29 which correspond to the framework regions (FRs or constant regions)of the light chain sequence of SEQ ID NO: 21 or the variable light chainsequence of SEQ ID NO: 22 or sequences that are at least 90% or 95%identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 2; the variable light chain region of SEQ IDNO: 22; the complementarity-determining regions (SEQ ID NO: 4; SEQ IDNO: 6; and SEQ ID NO: 8) of the variable heavy chain region of SEQ IDNO: 2; and the complementarity-determining regions (SEQ ID NO: 24; SEQID NO: 26; and SEQ ID NO: 28) of the variable light chain region of SEQID NO: 22 or sequences that are at least 90%, 95%, 96%, 97%, 98% or 99%identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 2; the variable light chain region of SEQ IDNO: 22; the framework regions (SEQ ID NO: 3; SEQ ID NO: 5; SEQ ID NO: 7;and SEQ ID NO: 9) of the variable heavy chain region of SEQ ID NO: 2;and the framework regions (SEQ ID NO: 23; SEQ ID NO: 25; SEQ ID NO: 27;and SEQ ID NO: 29) of the variable light chain region of SEQ ID NO: 22or sequences that are at least 90% or 95% identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab1, comprising, or alternatively consisting of, SEQ ID NO:1 and SEQ ID NO: 21, or an antibody or antibody fragment comprising theCDRs of Ab1 and having at least one of the biological activities setforth herein or is an anti-PCSK9 antibody that competes with Ab1 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab1 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab1.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab1, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 2 and the variable lightchain sequence of SEQ ID NO: 22 or sequences that are at least 90%, 95%,96%, 97%, 98% or 99% identical thereto. This embodiment of the inventionfurther includes Fabs containing additions, deletions, and variants ofSEQ ID NO: 2 and/or SEQ ID NO: 22 which retain the binding specificityfor PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab1.In another embodiment of the invention, anti-PCSK9 antibodies such asAb1 or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example haploid or diploid yeast suchas haploid or diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab1 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab2

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 41) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGLEWIGIIDAIDNTYYASWAKGRFTISKTSTTVDLKMTSLTTGDTATYFCARASILGYSIATGFNIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 42) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGLEWIGIIDAIDNTYYASWAKGRFTISKTSTTVDLKMTSLTTGDTATYFCARASILGY SIATGFNIWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab2 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 50) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 61) AYDMTQTPASVEVAVGGTVTIKCQASQSISSHLAWYQQKSGQPPKLLIYRASTLESGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYGVSDVDNGFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 62) AYDMTQTPASVEVAVGGTVTIKCQASQSISSHLAWYQQKSGQPPKLLIYRASTLESGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYGVSDVDNG FGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab2 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 70) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 44; SEQ ID NO: 46; andSEQ ID NO: 48 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 41 or which contain the variable heavy chain sequence of SEQID NO: 42, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 64; SEQ ID NO: 66; and SEQ ID NO: 68which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the light chain sequence of SEQ ID NO: 61 orwhich contain the variable light chain sequence of SEQ ID NO: 62, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 43; SEQ ID NO: 45; SEQ ID NO: 47; and SEQ ID NO:49 which correspond to the framework regions (FRs or constant regions)of the heavy chain sequence of SEQ ID NO: 41 or the variable heavy chainsequence of SEQ ID NO: 42, and/or one, two, three, or four of thepolypeptide sequences of SEQ ID NO: 63; SEQ ID NO: 65; SEQ ID NO: 67;and SEQ ID NO: 69 which correspond to the framework regions (FRs orconstant regions) of the light chain sequence of SEQ ID NO: 61 or thevariable light chain sequence of SEQ ID NO: 62, or combinations of thesepolypeptide sequences or sequences which are at least 80%, 90%, 95%,96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 41 or SEQ ID NO: 42 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 61 orSEQ ID NO: 62 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 44; SEQ ID NO: 46; and SEQ ID NO: 48 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 41 or the variable heavy chainsequence of SEQ ID NO: 42 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 64; SEQ ID NO: 66; and SEQ ID NO: 68 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 61 or the variable light chainsequence of SEQ ID NO: 62 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 43; SEQ ID NO: 45; SEQ ID NO: 47; and SEQ ID NO: 49 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 41 or the variable heavy chainsequence of SEQ ID NO: 42 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 63; SEQ ID NO: 65; SEQ ID NO: 67; and SEQ ID NO:69 which correspond to the framework regions (FRs or constant regions)of the light chain sequence of SEQ ID NO: 61 or the variable light chainsequence of SEQ ID NO: 62 or sequences that are at least 90% or 95%identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 42; the variable light chain region of SEQ IDNO: 62; the complementarity-determining regions (SEQ ID NO: 44; SEQ IDNO: 46; and SEQ ID NO: 48) of the variable heavy chain region of SEQ IDNO: 42; and the complementarity-determining regions (SEQ ID NO: 64; SEQID NO: 66; and SEQ ID NO: 68) of the variable light chain region of SEQID NO: 62 or sequences that are at least 90%, 95%, 96%, 97%, 98% or 99%identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 42; the variable light chain region of SEQ IDNO: 62; the framework regions (SEQ ID NO: 43; SEQ ID NO: 45; SEQ ID NO:47; and SEQ ID NO: 49) of the variable heavy chain region of SEQ ID NO:42; and the framework regions (SEQ ID NO: 63; SEQ ID NO: 65; SEQ ID NO:67; and SEQ ID NO: 69) of the variable light chain region of SEQ ID NO:62 or sequences that are at least 90% or 95% identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab2, comprising, or alternatively consisting of, SEQ ID NO:41 and SEQ ID NO: 61, or an antibody or antibody fragment comprising theCDRs of Ab2 and having at least one of the biological activities setforth herein or is an anti-PCSK9 antibody that competes with Ab2 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab2 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab2.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab2, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 42 and the variable lightchain sequence of SEQ ID NO: 62 or sequences that are at least 90%, 95%,96%, 97%, 98% or 99% identical thereto. This embodiment of the inventionfurther includes Fabs containing additions, deletions, and variants ofSEQ ID NO: 42 and/or SEQ ID NO: 62 which retain the binding specificityfor PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab2.In another embodiment of the invention, anti-PCSK9 antibodies such asAb2 or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example haploid or diploid yeast suchas haploid or diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab2 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab3

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 81) QSVEESGGRLVTPGGSLTLTCTASGFSLSSYYMSWVRQAPGKGLEWIGIIYPSGSTYYASWAKGRFTISKTSTTVDLKITSPTVEDTATYFCARGGAYATLNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 82) QSVEESGGRLVTPGGSLTLTCTASGFSLSSYYMSWVRQAPGKGLEWIGIIYPSGSTYYASWAKGRFTISKTSTTVDLKITSPTVEDTATYFCARGGAYAT LNLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab3 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 90) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 101) AVLTQTPSPVSAAVGGTVTISCQSSQSVYHNNLLSWYQQKPGQPPKLLIYDASKLTSGVSSRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDDDADNGFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 102) AVLTQTPSPVSAAVGGTVTISCQSSQSVYHNNLLSWYQQKPGQPPKLLIYDASKLTSGVSSRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDDDADNG FGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab3 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 110) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 84; SEQ ID NO: 86; andSEQ ID NO: 88 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 81 or which contain the variable heavy chain sequence of SEQID NO: 82, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 104; SEQ ID NO: 106; and SEQ ID NO:108 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 101or which contain the variable light chain sequence of SEQ ID NO: 102, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 83; SEQ ID NO: 85; SEQ ID NO: 87; and SEQ ID NO:89 which correspond to the framework regions (FRs or constant regions)of the heavy chain sequence of SEQ ID NO: 81 or the variable heavy chainsequence of SEQ ID NO: 82, and/or one, two, three, or four of thepolypeptide sequences of SEQ ID NO: 103; SEQ ID NO: 105; SEQ ID NO: 107;and SEQ ID NO: 109 which correspond to the framework regions (FRs orconstant regions) of the light chain sequence of SEQ ID NO: 101 or thevariable light chain sequence of SEQ ID NO: 102, or combinations ofthese polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 81 or SEQ ID NO: 82 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 101 orSEQ ID NO: 102 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 84; SEQ ID NO: 86; and SEQ ID NO: 88 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 81 or the variable heavy chainsequence of SEQ ID NO: 82 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 104; SEQ ID NO: 106; and SEQ ID NO: 108 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 101 or the variable light chainsequence of SEQ ID NO: 102 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 83; SEQ ID NO: 85; SEQ ID NO: 87; and SEQ ID NO: 89 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 81 or the variable heavy chainsequence of SEQ ID NO: 82 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 103; SEQ ID NO: 105; SEQ ID NO: 107; and SEQ IDNO: 109 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 101 or the variablelight chain sequence of SEQ ID NO: 102 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 82; the variable light chain region of SEQ IDNO: 102; the complementarity-determining regions (SEQ ID NO: 84; SEQ IDNO: 86; and SEQ ID NO: 88) of the variable heavy chain region of SEQ IDNO: 82; and the complementarity-determining regions (SEQ ID NO: 104; SEQID NO: 106; and SEQ ID NO: 108) of the variable light chain region ofSEQ ID NO: 102 or sequences that are at least 90%, 95%, 96%, 97%, 98% or99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 82; the variable light chain region of SEQ IDNO: 102; the framework regions (SEQ ID NO: 83; SEQ ID NO: 85; SEQ ID NO:87; and SEQ ID NO: 89) of the variable heavy chain region of SEQ ID NO:82; and the framework regions (SEQ ID NO: 103; SEQ ID NO: 105; SEQ IDNO: 107; and SEQ ID NO: 109) of the variable light chain region of SEQID NO: 102 or sequences that are at least 90% or 95% identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab3, comprising, or alternatively consisting of, SEQ ID NO:81 and SEQ ID NO: 101, or an antibody or antibody fragment comprisingthe CDRs of Ab3 and having at least one of the biological activities setforth herein or is an anti-PCSK9 antibody that competes with Ab3 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab3 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab3.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab3, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 82 and the variable lightchain sequence of SEQ ID NO: 102 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 82 and/or SEQ ID NO: 102 which retain the bindingspecificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab3.In another embodiment of the invention, anti-PCSK9 antibodies such asAb3 or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example haploid or diploid yeast suchas haploid or diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab3 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab4

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 121) QSVEESGGRLVTPGTPLTLTCTVSGIDLSSYAMIWVRQAPEKGLEYIGYIGGIDSTYYASWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCGRWSGTSGYNTIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 122) QSVEESGGRLVTPGTPLTLTCTVSGIDLSSYAMIWVRQAPEKGLEYIGYIGGIDSTYYASWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCGRWSGTSG YNTIWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab4 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 130) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 141) DVVMTQTPASVEAAVGGTVTIKCQASQSIYSNLAWYQQKPGQPPKLLIYGASNLASGVSSRFKGSRSGTEYTLTISDLECADAATYYCQCTGGGDSGNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 142) DVVMTQTPASVEAAVGGTVTIKCQASQSIYSNLAWYQQKPGQPPKLLIYGASNLASGVSSRFKGSRSGTEYTLTISDLECADAATYYCQCTGGGDSGNTF GGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab4 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 150) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 124; SEQ ID NO: 126;and SEQ ID NO: 128 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 121 or which contain the variable heavy chain sequence of SEQID NO: 122, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 144; SEQ ID NO: 146; and SEQ ID NO:148 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 141or which contain the variable light chain sequence of SEQ ID NO: 142, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 123; SEQ ID NO: 125; SEQ ID NO: 127; and SEQ IDNO: 129 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 121 or the variableheavy chain sequence of SEQ ID NO: 122, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 143; SEQ ID NO: 145; SEQ IDNO: 147; and SEQ ID NO: 149 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 141or the variable light chain sequence of SEQ ID NO: 142, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 121 or SEQ ID NO: 122 or polypeptidesthat are at least 90% or 95% identical thereto.

In another embodiment of the invention, antibody fragments of theinvention comprise, or alternatively consist of, the polypeptidesequence of SEQ ID NO: 141 or SEQ ID NO: 142 or polypeptides that are atleast 90%, 95%, 96%, 97%, 98% or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 124; SEQ ID NO: 126; and SEQ ID NO: 128 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 121 or the variable heavy chainsequence of SEQ ID NO: 122 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 144; SEQ ID NO: 146; and SEQ ID NO: 148 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 141 or the variable light chainsequence of SEQ ID NO: 142 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 123; SEQ ID NO: 125; SEQ ID NO: 127; and SEQ ID NO: 129 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 121 or the variable heavy chainsequence of SEQ ID NO: 122 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 143; SEQ ID NO: 145; SEQ ID NO: 147; and SEQ IDNO: 149 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 141 or the variablelight chain sequence of SEQ ID NO: 142 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 122; the variable light chain region of SEQID NO: 142; the complementarity-determining regions (SEQ ID NO: 124; SEQID NO: 126; and SEQ ID NO: 128) of the variable heavy chain region ofSEQ ID NO: 122; and the complementarity-determining regions (SEQ ID NO:144; SEQ ID NO: 146; and SEQ ID NO: 148) of the variable light chainregion of SEQ ID NO: 142 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 122; the variable light chain region of SEQID NO: 142; the framework regions (SEQ ID NO: 123; SEQ ID NO: 125; SEQID NO: 127; and SEQ ID NO: 129) of the variable heavy chain region ofSEQ ID NO: 122; and the framework regions (SEQ ID NO: 143; SEQ ID NO:145; SEQ ID NO: 147; and SEQ ID NO: 149) of the variable light chainregion of SEQ ID NO: 142 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab4, comprising, or alternatively consisting of, SEQ ID NO:121 and SEQ ID NO: 141, or an antibody or antibody fragment comprisingthe CDRs of Ab4 and having at least one of the biological activities setforth herein or is an anti-PCSK9 antibody that competes with Ab4 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab4 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab4.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab4, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 122 and the variable lightchain sequence of SEQ ID NO: 142 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 122 and/or SEQ ID NO: 142 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab4.In another embodiment of the invention, anti-PCSK9 antibodies such asAb4 or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example haploid or diploid yeast suchas haploid or diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab4 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab5

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 161) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGLEWIGIISNSGTTYYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARGIYWYWRVFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 162) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGLEWIGIISNSGTTYYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARGIYWYW RVFNLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab5 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 170) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 181) AVLTQTPSPVSAAVGGTVTINCQASQSVYNNLLSWYQQKPGQPPKWYDASNLASGVPDRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDDDADNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 182) AVLTQTPSPVSAAVGGTVTINCQASQSVYNNLLSWYQQKPGQPPKLLIYDASNLASGVPDRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDDDADNAF GGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab5 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 190) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 164; SEQ ID NO: 166;and SEQ ID NO: 168 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 161 or which contain the variable heavy chain sequence of SEQID NO: 162, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 184; SEQ ID NO: 186; and SEQ ID NO:188 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 181or which contain the variable light chain sequence of SEQ ID NO: 182, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 163; SEQ ID NO: 165; SEQ ID NO: 167; and SEQ IDNO: 169 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 161 or the variableheavy chain sequence of SEQ ID NO: 162, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 183; SEQ ID NO: 185; SEQ IDNO: 187; and SEQ ID NO: 189 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 181or the variable light chain sequence of SEQ ID NO: 182, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 161 or SEQ ID NO: 162 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 181 orSEQ ID NO: 182 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 164; SEQ ID NO: 166; and SEQ ID NO: 168 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 161 or the variable heavy chainsequence of SEQ ID NO: 162 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 184; SEQ ID NO: 186; and SEQ ID NO: 188 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 181 or the variable light chainsequence of SEQ ID NO: 182 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 163; SEQ ID NO: 165; SEQ ID NO: 167; and SEQ ID NO: 169 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 161 or the variable heavy chainsequence of SEQ ID NO: 162 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 183; SEQ ID NO: 185; SEQ ID NO: 187; and SEQ IDNO: 189 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 181 or the variablelight chain sequence of SEQ ID NO: 182 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 162; the variable light chain region of SEQID NO: 182; the complementarity-determining regions (SEQ ID NO: 164; SEQID NO: 166; and SEQ ID NO: 168) of the variable heavy chain region ofSEQ ID NO: 162; and the complementarity-determining regions (SEQ ID NO:184; SEQ ID NO: 186; and SEQ ID NO: 188) of the variable light chainregion of SEQ ID NO: 182 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 162; the variable light chain region of SEQID NO: 182; the framework regions (SEQ ID NO: 163; SEQ ID NO: 165; SEQID NO: 167; and SEQ ID NO: 169) of the variable heavy chain region ofSEQ ID NO: 162; and the framework regions (SEQ ID NO: 183; SEQ ID NO:185; SEQ ID NO: 187; and SEQ ID NO: 189) of the variable light chainregion of SEQ ID NO: 182 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab5, comprising, or alternatively consisting of, SEQ ID NO:161 and SEQ ID NO: 181, or an antibody or antibody fragment comprisingthe CDRs of Ab5 and having at least one of the biological activities setforth herein or is an anti-PCSK9 antibody that competes with Ab5 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab5 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab5.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab5, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 162 and the variable lightchain sequence of SEQ ID NO: 182 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 162 and/or SEQ ID NO: 182 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab5.In another embodiment of the invention, anti-PCSK9 antibodies such asAb5 or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example haploid or diploid yeast suchas haploid or diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab5 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab6

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 201) QEQLEESGGDLVKPEGSLTLTCTASGFSFSSNYWICWVRQAPGKGLEWIGCIRDGGGTYYASWAKGRLTISMTSSTTVTLQLNSLTAADTATYFCASDINDGWLGQFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 202) QEQLEESGGDLVKPEGSLTLTCTASGFSFSSNYWICWVRQAPGKGLEWIGCIRDGGGTYYASWAKGRLTISMTSSTTVTLQLNSLTAADTATYFCASDIN DGWLGQFNLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab6 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 210) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 221) ADIVMTQTPASVEVAVGGTVTIKCQASQSISAYLAWYQQKPGQPPKLLIYRAYTLASGVPSRFKGSGSGTQFTLTISDLECADAATYYCQSYYSVTTNTYGNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 222) ADIVMTQTPASVEVAVGGTVTIKCQASQSISAYLAWYQQKPGQPPKLLIYRAYTLASGVPSRFKGSGSGTQFTLTISDLECADAATYYCQSYYSVTTNTY GNTFGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab6 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 230) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 204; SEQ ID NO: 206;and SEQ ID NO: 208 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 201 or which contain the variable heavy chain sequence of SEQID NO: 202, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 224; SEQ ID NO: 226; and SEQ ID NO:228 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 221or which contain the variable light chain sequence of SEQ ID NO: 222, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 203; SEQ ID NO: 205; SEQ ID NO: 207; and SEQ IDNO: 209 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 201 or the variableheavy chain sequence of SEQ ID NO: 202, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 223; SEQ ID NO: 225; SEQ IDNO: 227; and SEQ ID NO: 229 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 221or the variable light chain sequence of SEQ ID NO: 222, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 201 or SEQ ID NO: 202 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 221 orSEQ ID NO: 222 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 204; SEQ ID NO: 206; and SEQ ID NO: 208 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 201 or the variable heavy chainsequence of SEQ ID NO: 202 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 224; SEQ ID NO: 226; and SEQ ID NO: 228 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 221 or the variable light chainsequence of SEQ ID NO: 222 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 203; SEQ ID NO: 205; SEQ ID NO: 207; and SEQ ID NO: 209 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 201 or the variable heavy chainsequence of SEQ ID NO: 202 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 223; SEQ ID NO: 225; SEQ ID NO: 227; and SEQ IDNO: 229 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 221 or the variablelight chain sequence of SEQ ID NO: 222 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 202; the variable light chain region of SEQID NO: 222; the complementarity-determining regions (SEQ ID NO: 204; SEQID NO: 206; and SEQ ID NO: 208) of the variable heavy chain region ofSEQ ID NO: 202; and the complementarity-determining regions (SEQ ID NO:224; SEQ ID NO: 226; and SEQ ID NO: 228) of the variable light chainregion of SEQ ID NO: 222 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 202; the variable light chain region of SEQID NO: 222; the framework regions (SEQ ID NO: 203; SEQ ID NO: 205; SEQID NO: 207; and SEQ ID NO: 209) of the variable heavy chain region ofSEQ ID NO: 202; and the framework regions (SEQ ID NO: 223; SEQ ID NO:225; SEQ ID NO: 227; and SEQ ID NO: 229) of the variable light chainregion of SEQ ID NO: 222 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab6, comprising, or alternatively consisting of, SEQ ID NO:201 and SEQ ID NO: 221, or an antibody or antibody fragment comprisingthe CDRs of Ab6 and having at least one of the biological activities setforth herein or is an anti-PCSK9 antibody that competes with Ab6 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab6 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab6.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab6, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 202 and the variable lightchain sequence of SEQ ID NO: 222 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 202 and/or SEQ ID NO: 222 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab6.In another embodiment of the invention, anti-PCSK9 antibodies such asAb6 or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example haploid or diploid yeast suchas haploid or diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab6 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab7

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 241) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNYWICWVRQAPGKGLEWIGCIRDGGGTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDINDGWLGQFNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 242) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNYWICWVRQAPGKGLEWIGCIRDGGGTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDINDGWLGQFNLWGQGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab7 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 250) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 261) ADIVMTQSPSSLSASVGDRVTIKCQASQSISAYLAWYQQKPGKVPKLLIYRAYTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYYSVTTNTYGNTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 262) ADIVMTQSPSSLSASVGDRVTIKCQASQSISAYLAWYQQKPGKVPKLLIYRAYTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYYSVTTNTY GNTFGGGTKVEIK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab7 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 270) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 244; SEQ ID NO: 246;and SEQ ID NO: 248 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 241 or which contain the variable heavy chain sequence of SEQID NO: 242, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 264; SEQ ID NO: 266; and SEQ ID NO:268 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 261or which contain the variable light chain sequence of SEQ ID NO: 262, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 243; SEQ ID NO: 245; SEQ ID NO: 247; and SEQ IDNO: 249 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 241 or the variableheavy chain sequence of SEQ ID NO: 242, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 263; SEQ ID NO: 265; SEQ IDNO: 267; and SEQ ID NO: 269 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 261or the variable light chain sequence of SEQ ID NO: 262, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 241 or SEQ ID NO: 242 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 261 orSEQ ID NO: 262 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 244; SEQ ID NO: 246; and SEQ ID NO: 248 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 241 or the variable heavy chainsequence of SEQ ID NO: 242 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 264; SEQ ID NO: 266; and SEQ ID NO: 268 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 261 or the variable light chainsequence of SEQ ID NO: 262 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 243; SEQ ID NO: 245; SEQ ID NO: 247; and SEQ ID NO: 249 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 241 or the variable heavy chainsequence of SEQ ID NO: 242 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 263; SEQ ID NO: 265; SEQ ID NO: 267; and SEQ IDNO: 269 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 261 or the variablelight chain sequence of SEQ ID NO: 262 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 242; the variable light chain region of SEQID NO: 262; the complementarity-determining regions (SEQ ID NO: 244; SEQID NO: 246; and SEQ ID NO: 248) of the variable heavy chain region ofSEQ ID NO: 242; and the complementarity-determining regions (SEQ ID NO:264; SEQ ID NO: 266; and SEQ ID NO: 268) of the variable light chainregion of SEQ ID NO: 262 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 242; the variable light chain region of SEQID NO: 262; the framework regions (SEQ ID NO: 243; SEQ ID NO: 245; SEQID NO: 247; and SEQ ID NO: 249) of the variable heavy chain region ofSEQ ID NO: 242; and the framework regions (SEQ ID NO: 263; SEQ ID NO:265; SEQ ID NO: 267; and SEQ ID NO: 269) of the variable light chainregion of SEQ ID NO: 262 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab7, comprising, or alternatively consisting of, SEQ ID NO:241 and SEQ ID NO: 261, or an antibody or antibody fragment comprisingthe CDRs of Ab7 and having at least one of the biological activities setforth herein or is an anti-PCSK9 antibody that competes with Ab7 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab7 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab7.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab7, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 242 and the variable lightchain sequence of SEQ ID NO: 262 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 242 and/or SEQ ID NO: 262 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab7.In another embodiment of the invention, anti-PCSK9 antibodies such asAb7 or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example haploid or diploid yeast suchas haploid or diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab7 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab8

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 281) QEQLVESGGGLVQPEGSLTLTCTASGFSFTSDYYMCWVRQAPGKGLEWIGCISTGDGSTYYASWAKGRFTISKPSSTTVTLQMTRLTAADTATYFCARDRYYSYAYGAYVYASDLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 282) QEQLVESGGGLVQPEGSLTLTCTASGFSFTSDYYMCWVRQAPGKGLEWIGCISTGDGSTYYASWAKGRFTISKPSSTTVTLQMTRLTAADTATYFCARDRYYSYAYGAYVYASDLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab8 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 290) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 301) ADIVMTQTPASVSEPVGGTVTINCQASESIRNYLSWYQQKPGQRPKLLIYGASTLASGVPSRFKGSGSGTDFTLTISDLECADAATYYCQSNYGISSRSYVNGFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 302) ADIVMTQTPASVSEPVGGTVTINCQASESIRNYLSWYQQKPGQRPKLLIYGASTLASGVPSRFKGSGSGTDFTLTISDLECADAATYYCQSNYGISSRSY VNGFGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab8 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 310) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 284; SEQ ID NO: 286;and SEQ ID NO: 288 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 281 or which contain the variable heavy chain sequence of SEQID NO: 282, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 304; SEQ ID NO: 306; and SEQ ID NO:308 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 301or which contain the variable light chain sequence of SEQ ID NO: 302, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 283; SEQ ID NO: 285; SEQ ID NO: 287; and SEQ IDNO: 289 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 281 or the variableheavy chain sequence of SEQ ID NO: 282, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 303; SEQ ID NO: 305; SEQ IDNO: 307; and SEQ ID NO: 309 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 301or the variable light chain sequence of SEQ ID NO: 302, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 281 or SEQ ID NO: 282 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 301 orSEQ ID NO: 302 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 284; SEQ ID NO: 286; and SEQ ID NO: 288 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 281 or the variable heavy chainsequence of SEQ ID NO: 282 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 304; SEQ ID NO: 306; and SEQ ID NO: 308 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 301 or the variable light chainsequence of SEQ ID NO: 302 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 283; SEQ ID NO: 285; SEQ ID NO: 287; and SEQ ID NO: 289 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 281 or the variable heavy chainsequence of SEQ ID NO: 282 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 303; SEQ ID NO: 305; SEQ ID NO: 307; and SEQ IDNO: 309 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 301 or the variablelight chain sequence of SEQ ID NO: 302 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 282; the variable light chain region of SEQID NO: 302; the complementarity-determining regions (SEQ ID NO: 284; SEQID NO: 286; and SEQ ID NO: 288) of the variable heavy chain region ofSEQ ID NO: 282; and the complementarity-determining regions (SEQ ID NO:304; SEQ ID NO: 306; and SEQ ID NO: 308) of the variable light chainregion of SEQ ID NO: 302 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 282; the variable light chain region of SEQID NO: 302; the framework regions (SEQ ID NO: 283; SEQ ID NO: 285; SEQID NO: 287; and SEQ ID NO: 289) of the variable heavy chain region ofSEQ ID NO: 282; and the framework regions (SEQ ID NO: 303; SEQ ID NO:305; SEQ ID NO: 307; and SEQ ID NO: 309) of the variable light chainregion of SEQ ID NO: 302 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab8, comprising, or alternatively consisting of, SEQ ID NO:281 and SEQ ID NO: 301, or an antibody or antibody fragment comprisingthe CDRs of Ab8 and having at least one of the biological activities setforth herein or is an anti-PCSK9 antibody that competes with Ab8 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab8 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab8.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab8, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 282 and the variable lightchain sequence of SEQ ID NO: 302 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 282 and/or SEQ ID NO: 302 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab8.In another embodiment of the invention, anti-PCSK9 antibodies such asAb8 or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example haploid or diploid yeast suchas haploid or diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab8 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab9

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 321) QSVEESGGRLVTPGTPLTLTCTVSGIDLSSYAMGWVRQAPGKGLEYIGIIVSYGPTYYASWAKGRFTISKTSTTVDLKITSPTAEDTATYFCARDLDANSSGYYGCFNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 322) QSVEESGGRLVTPGTPLTLTCTVSGIDLSSYAMGWVRQAPGKGLEYIGIIVSYGPTYYASWAKGRFTISKTSTTVDLKITSPTAEDTATYFCARDLDANS SGYYGCFNIWGQGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab9 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 330) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 341) AVVLTQTPASVSAAVGGTVTIKCQASQSISTALAWYQQKPGQPPKLLIYAASPLASGVSSRFKSSGSGTEFTLTISDLECADAATYYCQSYYGSSNIAFGGGTELEILRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 342) AVVLTQTPASVSAAVGGTVTIKCQASQSISTALAWYQQKPGQPPKLLIYAASPLASGVSSRFKSSGSGTEFTLTISDLECADAATYYCQSYYGSSNIAFG GGTELEIL.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab9 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 350) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 324; SEQ ID NO: 326;and SEQ ID NO: 328 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 321 or which contain the variable heavy chain sequence of SEQID NO: 322, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 344; SEQ ID NO: 346; and SEQ ID NO:348 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 341or which contain the variable light chain sequence of SEQ ID NO: 342, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 323; SEQ ID NO: 325; SEQ ID NO: 327; and SEQ IDNO: 329 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 321 or the variableheavy chain sequence of SEQ ID NO: 322, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 343; SEQ ID NO: 345; SEQ IDNO: 347; and SEQ ID NO: 349 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 341or the variable light chain sequence of SEQ ID NO: 342, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 321 or SEQ ID NO: 322 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 341 orSEQ ID NO: 342 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 324; SEQ ID NO: 326; and SEQ ID NO: 328 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 321 or the variable heavy chainsequence of SEQ ID NO: 322 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 344; SEQ ID NO: 346; and SEQ ID NO: 348 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 341 or the variable light chainsequence of SEQ ID NO: 342 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 323; SEQ ID NO: 325; SEQ ID NO: 327; and SEQ ID NO: 329 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 321 or the variable heavy chainsequence of SEQ ID NO: 322 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 343; SEQ ID NO: 345; SEQ ID NO: 347; and SEQ IDNO: 349 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 341 or the variablelight chain sequence of SEQ ID NO: 342 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 322; the variable light chain region of SEQID NO: 342; the complementarity-determining regions (SEQ ID NO: 324; SEQID NO: 326; and SEQ ID NO: 328) of the variable heavy chain region ofSEQ ID NO: 322; and the complementarity-determining regions (SEQ ID NO:344; SEQ ID NO: 346; and SEQ ID NO: 348) of the variable light chainregion of SEQ ID NO: 342 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 322; the variable light chain region of SEQID NO: 342; the framework regions (SEQ ID NO: 323; SEQ ID NO: 325; SEQID NO: 327; and SEQ ID NO: 329) of the variable heavy chain region ofSEQ ID NO: 322; and the framework regions (SEQ ID NO: 343; SEQ ID NO:345; SEQ ID NO: 347; and SEQ ID NO: 349) of the variable light chainregion of SEQ ID NO: 342 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab9, comprising, or alternatively consisting of, SEQ ID NO:321 and SEQ ID NO: 341, or an antibody or antibody fragment comprisingthe CDRs of Ab9 and having at least one of the biological activities setforth herein or is an anti-PCSK9 antibody that competes with Ab9 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab9 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab9.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab9, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 322 and the variable lightchain sequence of SEQ ID NO: 342 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 322 and/or SEQ ID NO: 342 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab9.In another embodiment of the invention, anti-PCSK9 antibodies such asAb9 or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example haploid or diploid yeast suchas haploid or diploid Pichia) and other yeast strains. Suitable Pichiaspecies include, but are not limited to, Pichia pastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab9 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab10

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 361) QEQLEESGGDLVKPEGSLTLTCTASGFSFSSSYWICWVRQAPGKGLEWIACIRAGGGNYYANWAKGRFTISRTSSTTVTLQMTSLTAADTATYFCASDINDGWLGQFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 362) QEQLEESGGDLVKPEGSLTLTCTASGFSFSSSYWICWVRQAPGKGLEWIACIRAGGGNYYANWAKGRFTISRTSSTTVTLQMTSLTAADTATYFCASDIN DGWLGQFNLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab10 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 370) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 381) ANIVMTQTPASVEAAVGGTVTIKCQASQSISNYLAWYQQKPGQPPKLLIYRTSTLASGVPSRFKGSGSGTQFTLTISDLECADAATYYCQSYYSVTTVAYGNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 382) ANIVMTQTPASVEAAVGGTVTIKCQASQSISNYLAWYQQKPGQPPKLLIYRTSTLASGVPSRFKGSGSGTQFTLTISDLECADAATYYCQSYYSVTTVAY GNTFGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab10 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 390) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 364; SEQ ID NO: 366;and SEQ ID NO: 368 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 361 or which contain the variable heavy chain sequence of SEQID NO: 362, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 384; SEQ ID NO: 386; and SEQ ID NO:388 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 381or which contain the variable light chain sequence of SEQ ID NO: 382, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 363; SEQ ID NO: 365; SEQ ID NO: 367; and SEQ IDNO: 369 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 361 or the variableheavy chain sequence of SEQ ID NO: 362, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 383; SEQ ID NO: 385; SEQ IDNO: 387; and SEQ ID NO: 389 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 381or the variable light chain sequence of SEQ ID NO: 382, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 361 or SEQ ID NO: 362 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 381 orSEQ ID NO: 382 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 364; SEQ ID NO: 366; and SEQ ID NO: 368 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 361 or the variable heavy chainsequence of SEQ ID NO: 362 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 384; SEQ ID NO: 386; and SEQ ID NO: 388 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 381 or the variable light chainsequence of SEQ ID NO: 382 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 363; SEQ ID NO: 365; SEQ ID NO: 367; and SEQ ID NO: 369 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 361 or the variable heavy chainsequence of SEQ ID NO: 362 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 383; SEQ ID NO: 385; SEQ ID NO: 387; and SEQ IDNO: 389 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 381 or the variablelight chain sequence of SEQ ID NO: 382 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 362; the variable light chain region of SEQID NO: 382; the complementarity-determining regions (SEQ ID NO: 364; SEQID NO: 366; and SEQ ID NO: 368) of the variable heavy chain region ofSEQ ID NO: 362; and the complementarity-determining regions (SEQ ID NO:384; SEQ ID NO: 386; and SEQ ID NO: 388) of the variable light chainregion of SEQ ID NO: 382 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 362; the variable light chain region of SEQID NO: 382; the framework regions (SEQ ID NO: 363; SEQ ID NO: 365; SEQID NO: 367; and SEQ ID NO: 369) of the variable heavy chain region ofSEQ ID NO: 362; and the framework regions (SEQ ID NO: 383; SEQ ID NO:385; SEQ ID NO: 387; and SEQ ID NO: 389) of the variable light chainregion of SEQ ID NO: 382 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab10, comprising, or alternatively consisting of, SEQ ID NO:361 and SEQ ID NO: 381, or an antibody or antibody fragment comprisingthe CDRs of Ab10 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab10 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab10 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab10.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab10, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 362 and the variable lightchain sequence of SEQ ID NO: 382 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 362 and/or SEQ ID NO: 382 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab10.In another embodiment of the invention, anti-PCSK9 antibodies such asAb10 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab10 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab11

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 401) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSYWICWVRQAPGKGLEWIACIRAGGGNYYANSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDINDGWLGQFNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 402) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSYWICWVRQAPGKGLEWIACIRAGGGNYYANSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDINDGWLGQFNLWGQGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab11 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 410) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 421) ANIVMTQSPSSLSASVGDRVTITCQASQSISNYLAWYQQKPGKVPKLLIYRTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYYSVTTVAYGNTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 422) ANIVMTQSPSSLSASVGDRVTITCQASQSISNYLAWYQQKPGKVPKLLIYRTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYYSVTTVAY GNTFGGGTKVEIK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab11 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 430) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 404; SEQ ID NO: 406;and SEQ ID NO: 408 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 401 or which contain the variable heavy chain sequence of SEQID NO: 402, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 424; SEQ ID NO: 426; and SEQ ID NO:428 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 421or which contain the variable light chain sequence of SEQ ID NO: 422, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 403; SEQ ID NO: 405; SEQ ID NO: 407; and SEQ IDNO: 409 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 401 or the variableheavy chain sequence of SEQ ID NO: 402, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 423; SEQ ID NO: 425; SEQ IDNO: 427; and SEQ ID NO: 429 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 421or the variable light chain sequence of SEQ ID NO: 422, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 401 or SEQ ID NO: 402 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 421 orSEQ ID NO: 422 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 404; SEQ ID NO: 406; and SEQ ID NO: 408 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 401 or the variable heavy chainsequence of SEQ ID NO: 402 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 424; SEQ ID NO: 426; and SEQ ID NO: 428 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 421 or the variable light chainsequence of SEQ ID NO: 422 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 403; SEQ ID NO: 405; SEQ ID NO: 407; and SEQ ID NO: 409 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 401 or the variable heavy chainsequence of SEQ ID NO: 402 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 423; SEQ ID NO: 425; SEQ ID NO: 427; and SEQ IDNO: 429 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 421 or the variablelight chain sequence of SEQ ID NO: 422 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 402; the variable light chain region of SEQID NO: 422; the complementarity-determining regions (SEQ ID NO: 404; SEQID NO: 406; and SEQ ID NO: 408) of the variable heavy chain region ofSEQ ID NO: 402; and the complementarity-determining regions (SEQ ID NO:424; SEQ ID NO: 426; and SEQ ID NO: 428) of the variable light chainregion of SEQ ID NO: 422 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 402; the variable light chain region of SEQID NO: 422; the framework regions (SEQ ID NO: 403; SEQ ID NO: 405; SEQID NO: 407; and SEQ ID NO: 409) of the variable heavy chain region ofSEQ ID NO: 402; and the framework regions (SEQ ID NO: 423; SEQ ID NO:425; SEQ ID NO: 427; and SEQ ID NO: 429) of the variable light chainregion of SEQ ID NO: 422 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab11, comprising, or alternatively consisting of, SEQ ID NO:401 and SEQ ID NO: 421, or an antibody or antibody fragment comprisingthe CDRs of Ab11 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab11 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab11 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab11.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab11, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 402 and the variable lightchain sequence of SEQ ID NO: 422 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 402 and/or SEQ ID NO: 422 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab11.In another embodiment of the invention, anti-PCSK9 antibodies such asAb11 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab11 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab12

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 441) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSYWICWVRQAPGKGLEWIACIRAGGGNYYANSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDINDGWLGQFNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 442) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSYWICWVRQAPGKGLEWIACIRAGGGNYYANSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDINDGWLGQFNLWGQGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab12 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 450) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 461) ANIVMTQSPSSLSASVGDRVTIKCQASQSISNYLAWYQQKPGKVPKLLIYRTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYYSVTTVAYGNTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 462) ANIVMTQSPSSLSASVGDRVTIKCQASQSISNYLAWYQQKPGKVPKLLIYRTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYYSVTTVAY GNTFGGGTKVEIK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab12 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 470) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 444; SEQ ID NO: 446;and SEQ ID NO: 448 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 441 or which contain the variable heavy chain sequence of SEQID NO: 442, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 464; SEQ ID NO: 466; and SEQ ID NO:468 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 461or which contain the variable light chain sequence of SEQ ID NO: 462, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 443; SEQ ID NO: 445; SEQ ID NO: 447; and SEQ IDNO: 449 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 441 or the variableheavy chain sequence of SEQ ID NO: 442, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 463; SEQ ID NO: 465; SEQ IDNO: 467; and SEQ ID NO: 469 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 461or the variable light chain sequence of SEQ ID NO: 462, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 441 or SEQ ID NO: 442 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 461 orSEQ ID NO: 462 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 444; SEQ ID NO: 446; and SEQ ID NO: 448 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 441 or the variable heavy chainsequence of SEQ ID NO: 442 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 464; SEQ ID NO: 466; and SEQ ID NO: 468 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 461 or the variable light chainsequence of SEQ ID NO: 462 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 443; SEQ ID NO: 445; SEQ ID NO: 447; and SEQ ID NO: 449 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 441 or the variable heavy chainsequence of SEQ ID NO: 442 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 463; SEQ ID NO: 465; SEQ ID NO: 467; and SEQ IDNO: 469 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 461 or the variablelight chain sequence of SEQ ID NO: 462 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 442; the variable light chain region of SEQID NO: 462; the complementarity-determining regions (SEQ ID NO: 444; SEQID NO: 446; and SEQ ID NO: 448) of the variable heavy chain region ofSEQ ID NO: 442; and the complementarity-determining regions (SEQ ID NO:464; SEQ ID NO: 466; and SEQ ID NO: 468) of the variable light chainregion of SEQ ID NO: 462 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 442; the variable light chain region of SEQID NO: 462; the framework regions (SEQ ID NO: 443; SEQ ID NO: 445; SEQID NO: 447; and SEQ ID NO: 449) of the variable heavy chain region ofSEQ ID NO: 442; and the framework regions (SEQ ID NO: 463; SEQ ID NO:465; SEQ ID NO: 467; and SEQ ID NO: 469) of the variable light chainregion of SEQ ID NO: 462 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab12, comprising, or alternatively consisting of, SEQ ID NO:441 and SEQ ID NO: 461, or an antibody or antibody fragment comprisingthe CDRs of Ab12 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab12 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab12 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab12.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab12, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 442 and the variable lightchain sequence of SEQ ID NO: 462 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 442 and/or SEQ ID NO: 462 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab12.In another embodiment of the invention, anti-PCSK9 antibodies such asAb12 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab12 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab13

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 481) QSVEESGGRLVTPGTPLTLTCTVSGIDLSTYGVGWVRQAPGKGLEYIGIISSSGSTYYASWAKGRFTISKTSSTTVDLKMTSLTTEDTATYFCARDWSSTTGYYGYFNMWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 482) QSVEESGGRLVTPGTPLTLTCTVSGIDLSTYGVGWVRQAPGKGLEYIGIISSSGSTYYASWAKGRFTISKTSSTTVDLKMTSLTTEDTATYFCARDWSST TGYYGYFNMWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab13 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 490) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 501) AFELTQTPSPVSAAVGGTVTIKCQASQSISTALAWYQQKPGQPPKLLIYGASNLESGVPSRFSGSGSGTQFTLTISDLECADAAIYYCQSSYGSSTLAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 502) AFELTQTPSPVSAAVGGTVTIKCQASQSISTALAWYQQKPGQPPKLLIYGASNLESGVPSRFSGSGSGTQFTLTISDLECADAAIYYCQSSYGSSTLA FGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab13 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 510) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 484; SEQ ID NO: 486;and SEQ ID NO: 488 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 481 or which contain the variable heavy chain sequence of SEQID NO: 482, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 504; SEQ ID NO: 506; and SEQ ID NO:508 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 501or which contain the variable light chain sequence of SEQ ID NO: 502, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 483; SEQ ID NO: 485; SEQ ID NO: 487; and SEQ IDNO: 489 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 481 or the variableheavy chain sequence of SEQ ID NO: 482, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 503; SEQ ID NO: 505; SEQ IDNO: 507; and SEQ ID NO: 509 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 501or the variable light chain sequence of SEQ ID NO: 502, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 481 or SEQ ID NO: 482 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 501 orSEQ ID NO: 502 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 484; SEQ ID NO: 486; and SEQ ID NO: 488 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 481 or the variable heavy chainsequence of SEQ ID NO: 482 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 504; SEQ ID NO: 506; and SEQ ID NO: 508 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 501 or the variable light chainsequence of SEQ ID NO: 502 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 483; SEQ ID NO: 485; SEQ ID NO: 487; and SEQ ID NO: 489 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 481 or the variable heavy chainsequence of SEQ ID NO: 482 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 503; SEQ ID NO: 505; SEQ ID NO: 507; and SEQ IDNO: 509 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 501 or the variablelight chain sequence of SEQ ID NO: 502 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 482; the variable light chain region of SEQID NO: 502; the complementarity-determining regions (SEQ ID NO: 484; SEQID NO: 486; and SEQ ID NO: 488) of the variable heavy chain region ofSEQ ID NO: 482; and the complementarity-determining regions (SEQ ID NO:504; SEQ ID NO: 506; and SEQ ID NO: 508) of the variable light chainregion of SEQ ID NO: 502 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 482; the variable light chain region of SEQID NO: 502; the framework regions (SEQ ID NO: 483; SEQ ID NO: 485; SEQID NO: 487; and SEQ ID NO: 489) of the variable heavy chain region ofSEQ ID NO: 482; and the framework regions (SEQ ID NO: 503; SEQ ID NO:505; SEQ ID NO: 507; and SEQ ID NO: 509) of the variable light chainregion of SEQ ID NO: 502 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab13, comprising, or alternatively consisting of, SEQ ID NO:481 and SEQ ID NO: 501, or an antibody or antibody fragment comprisingthe CDRs of Ab13 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab13 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab13 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab13.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab13, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 482 and the variable lightchain sequence of SEQ ID NO: 502 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 482 and/or SEQ ID NO: 502 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab13.In another embodiment of the invention, anti-PCSK9 antibodies such asAb13 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab13 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab14

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 521) QEQLEESGGDLVKPEGSLTLTCTGSGFSFSSIAYMCWIRQAPGKGLEWIGCIGSGSGNTYYANWAKGRFTISKSSSTTVTLQMTSLTAADTATYFCASDTNNGWLGQFNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 522) QEQLEESGGDLVKPEGSLTLTCTGSGFSFSSIAYMCWIRQAPGKGLEWIGCIGSGSGNTYYANWAKGRFTISKSSSTTVTLQMTSLTAADTATYFCASDTNNGWLGQFNLWGQGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab14 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 530) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 541) ADIVMTQTPASVSAAVGGTVTINCQASQSISSYLAWYQQKPGQPPKLLIYRASTLASGVPSRFKGSGSGTQFTLTISDLECADAATYYCQGYYSVTTNTYGNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 542) ADIVMTQTPASVSAAVGGTVTINCQASQSISSYLAWYQQKPGQPPKLLIYRASTLASGVPSRFKGSGSGTQFTLTISDLECADAATYYCQGYYSVTTN TYGNTFGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab14 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 550) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 524; SEQ ID NO: 526;and SEQ ID NO: 528 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 521 or which contain the variable heavy chain sequence of SEQID NO: 522, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 544; SEQ ID NO: 546; and SEQ ID NO:548 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 541or which contain the variable light chain sequence of SEQ ID NO: 542, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 523; SEQ ID NO: 525; SEQ ID NO: 527; and SEQ IDNO: 529 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 521 or the variableheavy chain sequence of SEQ ID NO: 522, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 543; SEQ ID NO: 545; SEQ IDNO: 547; and SEQ ID NO: 549 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 541or the variable light chain sequence of SEQ ID NO: 542, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 521 or SEQ ID NO: 522 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 541 orSEQ ID NO: 542 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 524; SEQ ID NO: 526; and SEQ ID NO: 528 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 521 or the variable heavy chainsequence of SEQ ID NO: 522 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 544; SEQ ID NO: 546; and SEQ ID NO: 548 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 541 or the variable light chainsequence of SEQ ID NO: 542 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 523; SEQ ID NO: 525; SEQ ID NO: 527; and SEQ ID NO: 529 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 521 or the variable heavy chainsequence of SEQ ID NO: 522 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 543; SEQ ID NO: 545; SEQ ID NO: 547; and SEQ IDNO: 549 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 541 or the variablelight chain sequence of SEQ ID NO: 542 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 522; the variable light chain region of SEQID NO: 542; the complementarity-determining regions (SEQ ID NO: 524; SEQID NO: 526; and SEQ ID NO: 528) of the variable heavy chain region ofSEQ ID NO: 522; and the complementarity-determining regions (SEQ ID NO:544; SEQ ID NO: 546; and SEQ ID NO: 548) of the variable light chainregion of SEQ ID NO: 542 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 522; the variable light chain region of SEQID NO: 542; the framework regions (SEQ ID NO: 523; SEQ ID NO: 525; SEQID NO: 527; and SEQ ID NO: 529) of the variable heavy chain region ofSEQ ID NO: 522; and the framework regions (SEQ ID NO: 543; SEQ ID NO:545; SEQ ID NO: 547; and SEQ ID NO: 549) of the variable light chainregion of SEQ ID NO: 542 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab14, comprising, or alternatively consisting of, SEQ ID NO:521 and SEQ ID NO: 541, or an antibody or antibody fragment comprisingthe CDRs of Ab14 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab14 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab14 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab14.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab14, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 522 and the variable lightchain sequence of SEQ ID NO: 542 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 522 and/or SEQ ID NO: 542 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab14.In another embodiment of the invention, anti-PCSK9 antibodies such asAb14 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab14 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab15

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 561) QEQLEESGGDLVKPEGSLTLTCTASGFSFSSSYWICWVRQAPGKGLEWIACIDAGNSGSTYYASWAKGRFTISKASSTTVTLQMTSLTAADTATYFCASDLNDGWLGQFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 562) QEQLEESGGDLVKPEGSLTLTCTASGFSFSSSYWICWVRQAPGKGLEWIACIDAGNSGSTYYASWAKGRFTISKASSTTVTLQMTSLTAADTATYFCASDLNDGWLGQFNLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab15 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 570) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 581) ANIVMTQTPSPVSGAVGGTVTIKCQASQSISDYLAWYQQKPGQPPKLLIYRASTLASGVPSRFRGSGSGTEYTLTITDLECADAATYYCQSYYSVTTNTYGNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 582) ANIVMTQTPSPVSGAVGGTVTIKCQASQSISDYLAWYQQKPGQPPKLLIYRASTLASGVPSRFRGSGSGTEYTLTITDLECADAATYYCQSYYSVTTN TYGNTFGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab15 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 590) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 564; SEQ ID NO: 566;and SEQ ID NO: 568 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 561 or which contain the variable heavy chain sequence of SEQID NO: 562, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 584; SEQ ID NO: 586; and SEQ ID NO:588 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 581or which contain the variable light chain sequence of SEQ ID NO: 582, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 563; SEQ ID NO: 565; SEQ ID NO: 567; and SEQ IDNO: 569 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 561 or the variableheavy chain sequence of SEQ ID NO: 562, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 583; SEQ ID NO: 585; SEQ IDNO: 587; and SEQ ID NO: 589 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 581or the variable light chain sequence of SEQ ID NO: 582, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 561 or SEQ ID NO: 562 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 581 orSEQ ID NO: 582 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 564; SEQ ID NO: 566; and SEQ ID NO: 568 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 561 or the variable heavy chainsequence of SEQ ID NO: 562 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 584; SEQ ID NO: 586; and SEQ ID NO: 588 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 581 or the variable light chainsequence of SEQ ID NO: 582 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 563; SEQ ID NO: 565; SEQ ID NO: 567; and SEQ ID NO: 569 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 561 or the variable heavy chainsequence of SEQ ID NO: 562 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 583; SEQ ID NO: 585; SEQ ID NO: 587; and SEQ IDNO: 589 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 581 or the variablelight chain sequence of SEQ ID NO: 582 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 562; the variable light chain region of SEQID NO: 582; the complementarity-determining regions (SEQ ID NO: 564; SEQID NO: 566; and SEQ ID NO: 568) of the variable heavy chain region ofSEQ ID NO: 562; and the complementarity-determining regions (SEQ ID NO:584; SEQ ID NO: 586; and SEQ ID NO: 588) of the variable light chainregion of SEQ ID NO: 582 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 562; the variable light chain region of SEQID NO: 582; the framework regions (SEQ ID NO: 563; SEQ ID NO: 565; SEQID NO: 567; and SEQ ID NO: 569) of the variable heavy chain region ofSEQ ID NO: 562; and the framework regions (SEQ ID NO: 583; SEQ ID NO:585; SEQ ID NO: 587; and SEQ ID NO: 589) of the variable light chainregion of SEQ ID NO: 582 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab15, comprising, or alternatively consisting of, SEQ ID NO:561 and SEQ ID NO: 581, or an antibody or antibody fragment comprisingthe CDRs of Ab15 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab15 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab15 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab15.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab15, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 562 and the variable lightchain sequence of SEQ ID NO: 582 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 562 and/or SEQ ID NO: 582 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab15.In another embodiment of the invention, anti-PCSK9 antibodies such asAb15 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab15 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab16

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 601) QEQLVESGGGLVQPEGSLTLTCTASGFSFSSDYWICWVRQAPGKGLEWIGCIRDGGGSYYANWAKGRLTISMTSSTTVGLKMTSLTAADTATYFCASDINDGWLGQFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 602) QEQLVESGGGLVQPEGSLTLTCTASGFSFSSDYWICWVRQAPGKGLEWIGCIRDGGGSYYANWAKGRLTISMTSSTTVGLKMTSLTAADTATYFCASDIN DGWLGQFNLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab16 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 610) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 621) ADIVMTQTPASVEAAVGGTVTIKCQASQSISSYLAWYQQKPGQPPKLLIYRASTLASGVPSRFSGSGSGTEFTLTISDLECADAATYYCQSYYSVTTVTYGNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 622) ADIVMTQTPASVEAAVGGTVTIKCQASQSISSYLAWYQQKPGQPPKLLIYRASTLASGVPSRFSGSGSGTEFTLTISDLECADAATYYCQSYYSVTTVTY GNTFGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab16 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 630) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 604; SEQ ID NO: 606;and SEQ ID NO: 608 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 601 or which contain the variable heavy chain sequence of SEQID NO: 602, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 624; SEQ ID NO: 626; and SEQ ID NO:628 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 621or which contain the variable light chain sequence of SEQ ID NO: 622, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 603; SEQ ID NO: 605; SEQ ID NO: 607; and SEQ IDNO: 609 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 601 or the variableheavy chain sequence of SEQ ID NO: 602, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 623; SEQ ID NO: 625; SEQ IDNO: 627; and SEQ ID NO: 629 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 621or the variable light chain sequence of SEQ ID NO: 622, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 601 or SEQ ID NO: 602 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 621 orSEQ ID NO: 622 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 604; SEQ ID NO: 606; and SEQ ID NO: 608 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 601 or the variable heavy chainsequence of SEQ ID NO: 602 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 624; SEQ ID NO: 626; and SEQ ID NO: 628 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 621 or the variable light chainsequence of SEQ ID NO: 622 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 603; SEQ ID NO: 605; SEQ ID NO: 607; and SEQ ID NO: 609 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 601 or the variable heavy chainsequence of SEQ ID NO: 602 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 623; SEQ ID NO: 625; SEQ ID NO: 627; and SEQ IDNO: 629 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 621 or the variablelight chain sequence of SEQ ID NO: 622 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 602; the variable light chain region of SEQID NO: 622; the complementarity-determining regions (SEQ ID NO: 604; SEQID NO: 606; and SEQ ID NO: 608) of the variable heavy chain region ofSEQ ID NO: 602; and the complementarity-determining regions (SEQ ID NO:624; SEQ ID NO: 626; and SEQ ID NO: 628) of the variable light chainregion of SEQ ID NO: 622 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 602; the variable light chain region of SEQID NO: 622; the framework regions (SEQ ID NO: 603; SEQ ID NO: 605; SEQID NO: 607; and SEQ ID NO: 609) of the variable heavy chain region ofSEQ ID NO: 602; and the framework regions (SEQ ID NO: 623; SEQ ID NO:625; SEQ ID NO: 627; and SEQ ID NO: 629) of the variable light chainregion of SEQ ID NO: 622 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab16, comprising, or alternatively consisting of, SEQ ID NO:601 and SEQ ID NO: 621, or an antibody or antibody fragment comprisingthe CDRs of Ab16 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab16 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab16 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab16.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab16, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 602 and the variable lightchain sequence of SEQ ID NO: 622 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 602 and/or SEQ ID NO: 622 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab16.In another embodiment of the invention, anti-PCSK9 antibodies such asAb16 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab16 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab17

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 641) QEQLEESGGDLVKPEGSLTLTCTASGFSFSSSYWICWVRQAPGKGLEWIGCIRPGSADYYASWAKGRFTISRASSSTVTLQMTSLTAADTATYFCASDINDGWLGQFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 642) QEQLEESGGDLVKPEGSLTLTCTASGFSFSSSYWICWVRQAPGKGLEWIGCIRPGSADYYASWAKGRFTISRASSSTVTLQMTSLTAADTATYFCASDIN DGWLGQFNLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab17 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 650) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 661) ADVVMTQTPASVEAAVGGTVTIKCQASLSIADYLAWYLQKPGQPPKLLIYRASTLASGVPSRFKGSGSGTEYTLTISDLECADAATYYCQSYYSVTTNTYGNTFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 662) ADVVMTQTPASVEAAVGGTVTIKCQASLSIADYLAWYLQKPGQPPKLLIYRASTLASGVPSRFKGSGSGTEYTLTISDLECADAATYYCQSYYSVTTNTY GNTFGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab17 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 670) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 644; SEQ ID NO: 646;and SEQ ID NO: 648 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 641 or which contain the variable heavy chain sequence of SEQID NO: 642, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 664; SEQ ID NO: 666; and SEQ ID NO:668 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 661or which contain the variable light chain sequence of SEQ ID NO: 662, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 643; SEQ ID NO: 645; SEQ ID NO: 647; and SEQ IDNO: 649 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 641 or the variableheavy chain sequence of SEQ ID NO: 642, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 663; SEQ ID NO: 665; SEQ IDNO: 667; and SEQ ID NO: 669 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 661or the variable light chain sequence of SEQ ID NO: 662, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 641 or SEQ ID NO: 642 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 661 orSEQ ID NO: 662 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 644; SEQ ID NO: 646; and SEQ ID NO: 648 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 641 or the variable heavy chainsequence of SEQ ID NO: 642 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 664; SEQ ID NO: 666; and SEQ ID NO: 668 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 661 or the variable light chainsequence of SEQ ID NO: 662 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 643; SEQ ID NO: 645; SEQ ID NO: 647; and SEQ ID NO: 649 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 641 or the variable heavy chainsequence of SEQ ID NO: 642 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 663; SEQ ID NO: 665; SEQ ID NO: 667; and SEQ IDNO: 669 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 661 or the variablelight chain sequence of SEQ ID NO: 662 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 642; the variable light chain region of SEQID NO: 662; the complementarity-determining regions (SEQ ID NO: 644; SEQID NO: 646; and SEQ ID NO: 648) of the variable heavy chain region ofSEQ ID NO: 642; and the complementarity-determining regions (SEQ ID NO:664; SEQ ID NO: 666; and SEQ ID NO: 668) of the variable light chainregion of SEQ ID NO: 662 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 642; the variable light chain region of SEQID NO: 662; the framework regions (SEQ ID NO: 643; SEQ ID NO: 645; SEQID NO: 647; and SEQ ID NO: 649) of the variable heavy chain region ofSEQ ID NO: 642; and the framework regions (SEQ ID NO: 663; SEQ ID NO:665; SEQ ID NO: 667; and SEQ ID NO: 669) of the variable light chainregion of SEQ ID NO: 662 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab17, comprising, or alternatively consisting of, SEQ ID NO:641 and SEQ ID NO: 661, or an antibody or antibody fragment comprisingthe CDRs of Ab17 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab17 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab17 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab17.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab17, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 642 and the variable lightchain sequence of SEQ ID NO: 662 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 642 and/or SEQ ID NO: 662 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab17.In another embodiment of the invention, anti-PCSK9 antibodies such asAb17 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab17 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab18

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 681) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNYWICWVRQAPGKGLEWIGCIRDGGGTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDINDGWLGQFNLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 682) EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNYWICWVRQAPGKGLEWIGCIRDGGGTYYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDINDGWLGQFNLWGQGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab18 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 690) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 701) ADIVMTQSPSSLSASVGDRVTIKCQASQSISAYLAWYQQKPGKVPKLLIYRAYTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYYSVTTNTYGNTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 702) ADIVMTQSPSSLSASVGDRVTIKCQASQSISAYLAWYQQKPGKVPKLLIYRAYTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQSYYSVTTNTY GNTFGGGTKVEIK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab18 which contain a constant light chain sequence comprising thesequence set forth below:RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 710).

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 684; SEQ ID NO: 686;and SEQ ID NO: 688 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 681 or which contain the variable heavy chain sequence of SEQID NO: 682, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 704; SEQ ID NO: 706; and SEQ ID NO:708 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 701or which contain the variable light chain sequence of SEQ ID NO: 702, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 683; SEQ ID NO: 685; SEQ ID NO: 687; and SEQ IDNO: 689 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 681 or the variableheavy chain sequence of SEQ ID NO: 682, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 703; SEQ ID NO: 705; SEQ IDNO: 707; and SEQ ID NO: 709 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 701or the variable light chain sequence of SEQ ID NO: 702, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 681 or SEQ ID NO: 682 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 701 orSEQ ID NO: 702 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 684; SEQ ID NO: 686; and SEQ ID NO: 688 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 681 or the variable heavy chainsequence of SEQ ID NO: 682 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 704; SEQ ID NO: 706; and SEQ ID NO: 708 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 701 or the variable light chainsequence of SEQ ID NO: 702 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 683; SEQ ID NO: 685; SEQ ID NO: 687; and SEQ ID NO: 689 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 681 or the variable heavy chainsequence of SEQ ID NO: 682 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 703; SEQ ID NO: 705; SEQ ID NO: 707; and SEQ IDNO: 709 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 701 or the variablelight chain sequence of SEQ ID NO: 702 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 682; the variable light chain region of SEQID NO: 702; the complementarity-determining regions (SEQ ID NO: 684; SEQID NO: 686; and SEQ ID NO: 688) of the variable heavy chain region ofSEQ ID NO: 682; and the complementarity-determining regions (SEQ ID NO:704; SEQ ID NO: 706; and SEQ ID NO: 708) of the variable light chainregion of SEQ ID NO: 702 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 682; the variable light chain region of SEQID NO: 702; the framework regions (SEQ ID NO: 683; SEQ ID NO: 685; SEQID NO: 687; and SEQ ID NO: 689) of the variable heavy chain region ofSEQ ID NO: 682; and the framework regions (SEQ ID NO: 703; SEQ ID NO:705; SEQ ID NO: 707; and SEQ ID NO: 709) of the variable light chainregion of SEQ ID NO: 702 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab18, comprising, or alternatively consisting of, SEQ ID NO:681 and SEQ ID NO: 701, or an antibody or antibody fragment comprisingthe CDRs of Ab18 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab18 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab18 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab18.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab18, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 682 and the variable lightchain sequence of SEQ ID NO: 702 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 682 and/or SEQ ID NO: 702 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab18.In another embodiment of the invention, anti-PCSK9 antibodies such asAb18 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab18 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab19

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 721) EVQLVESGGGLVQPGGSLRLSCAASGIDLSSYAMGWVRQAPGKGLEYIGIIVSYGPTYYASWAKGRFTISRDNSKNTVYLQMNSLRAEDTATYFCARDLDAQSSGYYGAFNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 722) EVQLVESGGGLVQPGGSLRLSCAASGIDLSSYAMGWVRQAPGKGLEYIGIIVSYGPTYYASWAKGRFTISRDNSKNTVYLQMNSLRAEDTATYFCARDLDAQSSGYYGAFNIWGQGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab19 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 730) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 741) DIQMTQSPSTLSASVGDRVTITCQASQSISTALAWYQQKPGKAPKLLIYAASPLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQSYYGSSNIAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 742) DIQMTQSPSTLSASVGDRVTITCQASQSISTALAWYQQKPGKAPKLLIYAASPLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQSYYGSSNIAFG GGTKVEIK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab19 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 750) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 724; SEQ ID NO: 726;and SEQ ID NO: 728 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 721 or which contain the variable heavy chain sequence of SEQID NO: 722, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 744; SEQ ID NO: 746; and SEQ ID NO:748 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 741or which contain the variable light chain sequence of SEQ ID NO: 742, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 723; SEQ ID NO: 725; SEQ ID NO: 727; and SEQ IDNO: 729 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 721 or the variableheavy chain sequence of SEQ ID NO: 722, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 743; SEQ ID NO: 745; SEQ IDNO: 747; and SEQ ID NO: 749 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 741or the variable light chain sequence of SEQ ID NO: 742, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 721 or SEQ ID NO: 722 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 741 orSEQ ID NO: 742 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 724; SEQ ID NO: 726; and SEQ ID NO: 728 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 721 or the variable heavy chainsequence of SEQ ID NO: 722 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 744; SEQ ID NO: 746; and SEQ ID NO: 748 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 741 or the variable light chainsequence of SEQ ID NO: 742 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 723; SEQ ID NO: 725; SEQ ID NO: 727; and SEQ ID NO: 729 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 721 or the variable heavy chainsequence of SEQ ID NO: 722 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 743; SEQ ID NO: 745; SEQ ID NO: 747; and SEQ IDNO: 749 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 741 or the variablelight chain sequence of SEQ ID NO: 742 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 722; the variable light chain region of SEQID NO: 742; the complementarity-determining regions (SEQ ID NO: 724; SEQID NO: 726; and SEQ ID NO: 728) of the variable heavy chain region ofSEQ ID NO: 722; and the complementarity-determining regions (SEQ ID NO:744; SEQ ID NO: 746; and SEQ ID NO: 748) of the variable light chainregion of SEQ ID NO: 742 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 722; the variable light chain region of SEQID NO: 742; the framework regions (SEQ ID NO: 723; SEQ ID NO: 725; SEQID NO: 727; and SEQ ID NO: 729) of the variable heavy chain region ofSEQ ID NO: 722; and the framework regions (SEQ ID NO: 743; SEQ ID NO:745; SEQ ID NO: 747; and SEQ ID NO: 749) of the variable light chainregion of SEQ ID NO: 742 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab19, comprising, or alternatively consisting of, SEQ ID NO:721 and SEQ ID NO: 741, or an antibody or antibody fragment comprisingthe CDRs of Ab19 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab19 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab19 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab19.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab19, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 722 and the variable lightchain sequence of SEQ ID NO: 742 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 722 and/or SEQ ID NO: 742 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab19.In another embodiment of the invention, anti-PCSK9 antibodies such asAb19 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab19 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab20

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 761) EVQLVESGGGLVQPGGSLRLSCAASGIDLSSYAMGWVRQAPGKGLEYIGIIVSYGPTYYASWAKGRFTISRDNSKSTVYLQMNSLRAEDTATYFCARDLDAQSSGYYGAFNIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 762) EVQLVESGGGLVQPGGSLRLSCAASGIDLSSYAMGWVRQAPGKGLEYIGIIVSYGPTYYASWAKGRFTISRDNSKSTVYLQMNSLRAEDTATYFCARDLDAQSSGYYGAFNIWGQGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab20 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 770) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 781) DIQMTQSPSTLSASVGDRVTITCQASQSISTALAWYQQKPGKAPKLLIYAASPLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQSYYGSSNIAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 782) DIQMTQSPSTLSASVGDRVTITCQASQSISTALAWYQQKPGKAPKLLIYAASPLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQSYYGSSNIAFG GGTKVEIK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab20 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 790) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 764; SEQ ID NO: 766;and SEQ ID NO: 768 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 761 or which contain the variable heavy chain sequence of SEQID NO: 762, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 784; SEQ ID NO: 786; and SEQ ID NO:788 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 781or which contain the variable light chain sequence of SEQ ID NO: 782, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 763; SEQ ID NO: 765; SEQ ID NO: 767; and SEQ IDNO: 769 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 761 or the variableheavy chain sequence of SEQ ID NO: 762, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 783; SEQ ID NO: 785; SEQ IDNO: 787; and SEQ ID NO: 789 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 781or the variable light chain sequence of SEQ ID NO: 782, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 761 or SEQ ID NO: 762 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 781 orSEQ ID NO: 782 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 764; SEQ ID NO: 766; and SEQ ID NO: 768 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 761 or the variable heavy chainsequence of SEQ ID NO: 762 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 784; SEQ ID NO: 786; and SEQ ID NO: 788 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 781 or the variable light chainsequence of SEQ ID NO: 782 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 763; SEQ ID NO: 765; SEQ ID NO: 767; and SEQ ID NO: 769 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 761 or the variable heavy chainsequence of SEQ ID NO: 762 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 783; SEQ ID NO: 785; SEQ ID NO: 787; and SEQ IDNO: 789 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 781 or the variablelight chain sequence of SEQ ID NO: 782 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 762; the variable light chain region of SEQID NO: 782; the complementarity-determining regions (SEQ ID NO: 764; SEQID NO: 766; and SEQ ID NO: 768) of the variable heavy chain region ofSEQ ID NO: 762; and the complementarity-determining regions (SEQ ID NO:784; SEQ ID NO: 786; and SEQ ID NO: 788) of the variable light chainregion of SEQ ID NO: 782 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 762; the variable light chain region of SEQID NO: 782; the framework regions (SEQ ID NO: 763; SEQ ID NO: 765; SEQID NO: 767; and SEQ ID NO: 769) of the variable heavy chain region ofSEQ ID NO: 762; and the framework regions (SEQ ID NO: 783; SEQ ID NO:785; SEQ ID NO: 787; and SEQ ID NO: 789) of the variable light chainregion of SEQ ID NO: 782 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab20, comprising, or alternatively consisting of, SEQ ID NO:761 and SEQ ID NO: 781, or an antibody or antibody fragment comprisingthe CDRs of Ab20 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab20 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab20 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab20.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab20, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 762 and the variable lightchain sequence of SEQ ID NO: 782 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 762 and/or SEQ ID NO: 782 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab20.In another embodiment of the invention, anti-PCSK9 antibodies such asAb20 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab20 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab21

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 801) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMNWVRQAPGKGLEWIGAIRSSGATFFASWVNGRFTISKTSTTVDLKITSPTPEDTATYFCARDTNDGWYINRLDLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 802) QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMNWVRQAPGKGLEWIGAIRSSGATFFASWVNGRFTISKTSTTVDLKITSPTPEDTATYFCARDTNDGW YINRLDLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab21 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 810) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 821) AAVLTQTPSPVSAAVGGTVSISCQSSKSVYSNYLSWFQQKPGQPPKFLIYKASTLASGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAGGDTNISDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 822) AAVLTQTPSPVSAAVGGTVSISCQSSKSVYSNYLSWFQQKPGQPPKFLIYKASTLASGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAGGDTNISDNA FGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab21 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 830) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 804; SEQ ID NO: 806;and SEQ ID NO: 808 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 801 or which contain the variable heavy chain sequence of SEQID NO: 802, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 824; SEQ ID NO: 826; and SEQ ID NO:828 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 821or which contain the variable light chain sequence of SEQ ID NO: 822, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 803; SEQ ID NO: 805; SEQ ID NO: 807; and SEQ IDNO: 809 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 801 or the variableheavy chain sequence of SEQ ID NO: 802, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 823; SEQ ID NO: 825; SEQ IDNO: 827; and SEQ ID NO: 829 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 821or the variable light chain sequence of SEQ ID NO: 822, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 801 or SEQ ID NO: 802 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 821 orSEQ ID NO: 822 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 804; SEQ ID NO: 806; and SEQ ID NO: 808 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 801 or the variable heavy chainsequence of SEQ ID NO: 802 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 824; SEQ ID NO: 826; and SEQ ID NO: 828 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 821 or the variable light chainsequence of SEQ ID NO: 822 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 803; SEQ ID NO: 805; SEQ ID NO: 807; and SEQ ID NO: 809 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 801 or the variable heavy chainsequence of SEQ ID NO: 802 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 823; SEQ ID NO: 825; SEQ ID NO: 827; and SEQ IDNO: 829 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 821 or the variablelight chain sequence of SEQ ID NO: 822 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 802; the variable light chain region of SEQID NO: 822; the complementarity-determining regions (SEQ ID NO: 804; SEQID NO: 806; and SEQ ID NO: 808) of the variable heavy chain region ofSEQ ID NO: 802; and the complementarity-determining regions (SEQ ID NO:824; SEQ ID NO: 826; and SEQ ID NO: 828) of the variable light chainregion of SEQ ID NO: 822 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 802; the variable light chain region of SEQID NO: 822; the framework regions (SEQ ID NO: 803; SEQ ID NO: 805; SEQID NO: 807; and SEQ ID NO: 809) of the variable heavy chain region ofSEQ ID NO: 802; and the framework regions (SEQ ID NO: 823; SEQ ID NO:825; SEQ ID NO: 827; and SEQ ID NO: 829) of the variable light chainregion of SEQ ID NO: 822 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab21, comprising, or alternatively consisting of, SEQ ID NO:801 and SEQ ID NO: 821, or an antibody or antibody fragment comprisingthe CDRs of Ab21 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab21 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab21 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab21.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab21, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 802 and the variable lightchain sequence of SEQ ID NO: 822 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 802 and/or SEQ ID NO: 822 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab21.In another embodiment of the invention, anti-PCSK9 antibodies such asAb21 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab21 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab22

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 841) EVQLVESGGGLVQPGGSLRLSCAASGFSLSSYAMNWVRQAPGKGLEWIGAIRSSGATFFASSVNGRFTISRDNSKNTVYLQMNSLRAEDTAVYYCARDTNDGWYINRLDLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 842) EVQLVESGGGLVQPGGSLRLSCAASGFSLSSYAMNWVRQAPGKGLEWIGAIRSSGATFFASSVNGRFTISRDNSKNTVYLQMNSLRAEDTAVYYCARDTNDGWYINRLDLWGQGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab22 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 850) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 861) AVLTQSPSTLSASVGDRVTITCQSSKSVYSNYLSWFQQKPGKAPKFLIYKASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCAGGDTNIADNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 862) AVLTQSPSTLSASVGDRVTITCQSSKSVYSNYLSWFQQKPGKAPKFLIYKASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCAGGDTNIADNAF GGGTKVEIK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab22 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 870) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 844; SEQ ID NO: 846;and SEQ ID NO: 848 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 841 or which contain the variable heavy chain sequence of SEQID NO: 842, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 864; SEQ ID NO: 866; and SEQ ID NO:868 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 861or which contain the variable light chain sequence of SEQ ID NO: 862, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 843; SEQ ID NO: 845; SEQ ID NO: 847; and SEQ IDNO: 849 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 841 or the variableheavy chain sequence of SEQ ID NO: 842, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 863; SEQ ID NO: 865; SEQ IDNO: 867; and SEQ ID NO: 869 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 861or the variable light chain sequence of SEQ ID NO: 862, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 841 or SEQ ID NO: 842 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 861 orSEQ ID NO: 862 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 844; SEQ ID NO: 846; and SEQ ID NO: 848 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 841 or the variable heavy chainsequence of SEQ ID NO: 842 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 864; SEQ ID NO: 866; and SEQ ID NO: 868 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 861 or the variable light chainsequence of SEQ ID NO: 862 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 843; SEQ ID NO: 845; SEQ ID NO: 847; and SEQ ID NO: 849 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 841 or the variable heavy chainsequence of SEQ ID NO: 842 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 863; SEQ ID NO: 865; SEQ ID NO: 867; and SEQ IDNO: 869 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 861 or the variablelight chain sequence of SEQ ID NO: 862 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 842; the variable light chain region of SEQID NO: 862; the complementarity-determining regions (SEQ ID NO: 844; SEQID NO: 846; and SEQ ID NO: 848) of the variable heavy chain region ofSEQ ID NO: 842; and the complementarity-determining regions (SEQ ID NO:864; SEQ ID NO: 866; and SEQ ID NO: 868) of the variable light chainregion of SEQ ID NO: 862 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 842; the variable light chain region of SEQID NO: 862; the framework regions (SEQ ID NO: 843; SEQ ID NO: 845; SEQID NO: 847; and SEQ ID NO: 849) of the variable heavy chain region ofSEQ ID NO: 842; and the framework regions (SEQ ID NO: 863; SEQ ID NO:865; SEQ ID NO: 867; and SEQ ID NO: 869) of the variable light chainregion of SEQ ID NO: 862 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab22, comprising, or alternatively consisting of, SEQ ID NO:841 and SEQ ID NO: 861, or an antibody or antibody fragment comprisingthe CDRs of Ab22 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab22 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab22 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab22.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab22, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 842 and the variable lightchain sequence of SEQ ID NO: 862 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 842 and/or SEQ ID NO: 862 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab22.In another embodiment of the invention, anti-PCSK9 antibodies such asAb22 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab22 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab23

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 881) QSLEESGGDLVKPGASLTLTCKASGFSFSSGYYMCWVRQAPGKGLEWIACIYAGSGGSTFFANWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCARDGGYAGYGYAFFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 882) QSLEESGGDLVKPGASLTLTCKASGFSFSSGYYMCWVRQAPGKGLEWIACIYAGSGGSTFFANWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCARDGGYAGYGYAFFNLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab23 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 890) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 901) DVVMTQTPASVSEPVGGTVTIKCQASERIYSGLAWYQQKPGQPPKLLIYGASTLASGVPSRFKGSGSGTDFTLTISDLECDDAAIYYCQCTYYGSSYPNVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 902) DVVMTQTPASVSEPVGGTVTIKCQASERIYSGLAWYQQKPGQPPKLLIYGASTLASGVPSRFKGSGSGTDFTLTISDLECDDAAIYYCQCTYYGSSYPNV FGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab23 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 910) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 884; SEQ ID NO: 886;and SEQ ID NO: 888 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 881 or which contain the variable heavy chain sequence of SEQID NO: 882, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 904; SEQ ID NO: 906; and SEQ ID NO:908 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 901or which contain the variable light chain sequence of SEQ ID NO: 902, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 883; SEQ ID NO: 885; SEQ ID NO: 887; and SEQ IDNO: 889 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 881 or the variableheavy chain sequence of SEQ ID NO: 882, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 903; SEQ ID NO: 905; SEQ IDNO: 907; and SEQ ID NO: 909 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 901or the variable light chain sequence of SEQ ID NO: 902, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 881 or SEQ ID NO: 882 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 901 orSEQ ID NO: 902 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 884; SEQ ID NO: 886; and SEQ ID NO: 888 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 881 or the variable heavy chainsequence of SEQ ID NO: 882 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 904; SEQ ID NO: 906; and SEQ ID NO: 908 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 901 or the variable light chainsequence of SEQ ID NO: 902 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 883; SEQ ID NO: 885; SEQ ID NO: 887; and SEQ ID NO: 889 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 881 or the variable heavy chainsequence of SEQ ID NO: 882 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 903; SEQ ID NO: 905; SEQ ID NO: 907; and SEQ IDNO: 909 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 901 or the variablelight chain sequence of SEQ ID NO: 902 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 882; the variable light chain region of SEQID NO: 902; the complementarity-determining regions (SEQ ID NO: 884; SEQID NO: 886; and SEQ ID NO: 888) of the variable heavy chain region ofSEQ ID NO: 882; and the complementarity-determining regions (SEQ ID NO:904; SEQ ID NO: 906; and SEQ ID NO: 908) of the variable light chainregion of SEQ ID NO: 902 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 882; the variable light chain region of SEQID NO: 902; the framework regions (SEQ ID NO: 883; SEQ ID NO: 885; SEQID NO: 887; and SEQ ID NO: 889) of the variable heavy chain region ofSEQ ID NO: 882; and the framework regions (SEQ ID NO: 903; SEQ ID NO:905; SEQ ID NO: 907; and SEQ ID NO: 909) of the variable light chainregion of SEQ ID NO: 902 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab23, comprising, or alternatively consisting of, SEQ ID NO:881 and SEQ ID NO: 901, or an antibody or antibody fragment comprisingthe CDRs of Ab23 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab23 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab23 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab23.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab23, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 882 and the variable lightchain sequence of SEQ ID NO: 902 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 882 and/or SEQ ID NO: 902 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab23.In another embodiment of the invention, anti-PCSK9 antibodies such asAb23 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab23 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

Antibody Ab24

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess a heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 921) QSLEESGGDLVKPGASLTLTCKASGFSFSSGYYMCWVRQAPGKGLEWIACIYAGSGGSTFFANWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCARDGGYAGYGYAFFNLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK.

In one embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variableheavy chain sequence comprising the sequence set forth below:

(SEQ ID NO: 922) QSLEESGGDLVKPGASLTLTCKASGFSFSSGYYMCWVRQAPGKGLEWIACIYAGSGGSTFFANWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCARDGGYAGYGYAFFNLWGPGTLVTVSS.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that possess the sameepitopic specificity as Ab24 and which contain a constant heavy chainsequence comprising the sequence set forth below:

(SEQ ID NO: 930) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 941) DVVMTQTPASVSEPVGGTVTIKCQASERIYSGLAWYQQKPGQPPKLLIYGASTLASGVPSRFKGSGSGTDFTLTISDLECDDAAIYYCQATYYGSSYPNVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYA CEVTHQGLSSPVTKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain a variablelight chain sequence comprising the sequence set forth below:

(SEQ ID NO: 942) DVVMTQTPASVSEPVGGTVTIKCQASERIYSGLAWYQQKPGQPPKLLIYGASTLASGVPSRFKGSGSGTDFTLTISDLECDDAAIYYCQATYYGSSYP NVFGGGTEVVVK.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that bind the same epitopeas Ab24 which contain a constant light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 950) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC.

In another embodiment, the invention includes antibodies and antibodyfragments having binding specificity to PCSK9 that contain one, two, orthree of the polypeptide sequences of SEQ ID NO: 924; SEQ ID NO: 926;and SEQ ID NO: 928 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the heavy chain sequence ofSEQ ID NO: 921 or which contain the variable heavy chain sequence of SEQID NO: 922, and/or which further contain one, two, or three of thepolypeptide sequences of SEQ ID NO: 944; SEQ ID NO: 946; and SEQ ID NO:948 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain sequence of SEQ ID NO: 941or which contain the variable light chain sequence of SEQ ID NO: 942, orantibodies or fragments containing combinations of sequences which areat least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical thereto. Inanother embodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the exemplified variable heavy chain and variable lightchain sequences, or the heavy chain and light chain sequences set forthabove, or sequences that are at least 90% or 95% identical thereto.

The invention further contemplates anti-PCSK9 antibodies and antibodyfragments comprising one, two, three, or four of the polypeptidesequences of SEQ ID NO: 923; SEQ ID NO: 925; SEQ ID NO: 927; and SEQ IDNO: 929 which correspond to the framework regions (FRs or constantregions) of the heavy chain sequence of SEQ ID NO: 921 or the variableheavy chain sequence of SEQ ID NO: 922, and/or one, two, three, or fourof the polypeptide sequences of SEQ ID NO: 943; SEQ ID NO: 945; SEQ IDNO: 947; and SEQ ID NO: 949 which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 941or the variable light chain sequence of SEQ ID NO: 942, or combinationsof these polypeptide sequences or sequences which are at least 80%, 90%,95%, 96%, 97%, 98% or 99% identical therewith.

In another embodiment of the invention, the antibodies and antibodyfragments of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the FRs, CDRs,the variable heavy chain and variable light chain sequences, and theheavy chain and light chain sequences set forth above, including all ofthem or sequences which are at least 90% or 95% identical thereto.

In another embodiment of the invention, the anti-PCSK9 antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 921 or SEQ ID NO: 922 or polypeptidesthat are at least 90% or 95% identical thereto. In another embodiment ofthe invention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 941 orSEQ ID NO: 942 or polypeptides that are at least 90%, 95%, 96%, 97%, 98%or 99% identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 924; SEQ ID NO: 926; and SEQ ID NO: 928 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe heavy chain sequence of SEQ ID NO: 921 or the variable heavy chainsequence of SEQ ID NO: 922 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, or three of the polypeptide sequences of SEQ IDNO: 944; SEQ ID NO: 946; and SEQ ID NO: 948 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe light chain sequence of SEQ ID NO: 941 or the variable light chainsequence of SEQ ID NO: 942 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the antibody or antibodyfragment having binding specificity to PCSK9 comprises, or alternativelyconsists of, one, two, three, or four of the polypeptide sequences ofSEQ ID NO: 923; SEQ ID NO: 925; SEQ ID NO: 927; and SEQ ID NO: 929 whichcorrespond to the framework regions (FRs or constant regions) of theheavy chain sequence of SEQ ID NO: 921 or the variable heavy chainsequence of SEQ ID NO: 922 or sequences that are at least 90% or 95%identical thereto.

In a further embodiment of the invention, the subject antibody orantibody fragment having binding specificity to PCSK9 comprises, oralternatively consists of, one, two, three, or four of the polypeptidesequences of SEQ ID NO: 943; SEQ ID NO: 945; SEQ ID NO: 947; and SEQ IDNO: 949 which correspond to the framework regions (FRs or constantregions) of the light chain sequence of SEQ ID NO: 941 or the variablelight chain sequence of SEQ ID NO: 942 or sequences that are at least90% or 95% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 922; the variable light chain region of SEQID NO: 942; the complementarity-determining regions (SEQ ID NO: 924; SEQID NO: 926; and SEQ ID NO: 928) of the variable heavy chain region ofSEQ ID NO: 922; and the complementarity-determining regions (SEQ ID NO:944; SEQ ID NO: 946; and SEQ ID NO: 948) of the variable light chainregion of SEQ ID NO: 942 or sequences that are at least 90%, 95%, 96%,97%, 98% or 99% identical thereto.

The invention also contemplates antibody fragments that include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toPCSK9 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable heavychain region of SEQ ID NO: 922; the variable light chain region of SEQID NO: 942; the framework regions (SEQ ID NO: 923; SEQ ID NO: 925; SEQID NO: 927; and SEQ ID NO: 929) of the variable heavy chain region ofSEQ ID NO: 922; and the framework regions (SEQ ID NO: 943; SEQ ID NO:945; SEQ ID NO: 947; and SEQ ID NO: 949) of the variable light chainregion of SEQ ID NO: 942 or sequences that are at least 90% or 95%identical thereto.

In a particularly preferred embodiment of the invention, the anti-PCSK9antibody is Ab24, comprising, or alternatively consisting of, SEQ ID NO:921 and SEQ ID NO: 941, or an antibody or antibody fragment comprisingthe CDRs of Ab24 and having at least one of the biological activitiesset forth herein or is an anti-PCSK9 antibody that competes with Ab24 inbinding PCSK9, preferably one containing sequences that are at least90%, 95%, 96%, 97%, 98% or 99% identical to that of Ab24 or an antibodythat binds to the same or overlapping epitope(s) on PCSK9 as Ab24.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for PCSK9. Withrespect to antibody Ab24, the Fab fragment preferably includes thevariable heavy chain sequence of SEQ ID NO: 922 and the variable lightchain sequence of SEQ ID NO: 942 or sequences that are at least 90%,95%, 96%, 97%, 98% or 99% identical thereto. This embodiment of theinvention further includes Fabs containing additions, deletions, andvariants of SEQ ID NO: 922 and/or SEQ ID NO: 942 which retain thebinding specificity for PCSK9.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab24.In another embodiment of the invention, anti-PCSK9 antibodies such asAb24 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example haploid or diploidyeast such as haploid or diploid Pichia) and other yeast strains.Suitable Pichia species include, but are not limited to, Pichiapastoris.

In an additional embodiment, the invention is further directed topolynucleotides encoding antibody polypeptides having bindingspecificity to PCSK9, including the heavy and/or light chains of Ab24 aswell as fragments, variants, combinations of one or more of the FRs,CDRs, the variable heavy chain and variable light chain sequences, andthe heavy chain and light chain sequences set forth above, including allof them or sequences which are at least 90% or 95% identical thereto.

In another embodiment, the invention contemplates an isolated anti-PCSK9antibody comprising a V_(H) polypeptide sequence selected from: SEQ IDNO: 2, SEQ ID NO: 42, SEQ ID NO: 82, SEQ ID NO: 122, SEQ ID NO: 162, SEQID NO: 202, SEQ ID NO: 242, SEQ ID NO: 282, SEQ ID NO: 322, SEQ ID NO:362, SEQ ID NO: 402, SEQ ID NO: 442, SEQ ID NO: 482, SEQ ID NO: 522, SEQID NO: 562, SEQ ID NO: 602, SEQ ID NO: 642, SEQ ID NO: 692, SEQ ID NO:732, SEQ ID NO: 772, SEQ ID NO: 812, SEQ ID NO: 852, SEQ ID NO: 892, SEQID NO: 932, or a variant thereof; and further comprising a V_(L)polypeptide sequence selected from: SEQ ID NO: 22, SEQ ID NO: 62, SEQ IDNO: 102, SEQ ID NO: 142, SEQ ID NO: 182, SEQ ID NO: 222, SEQ ID NO: 262,SEQ ID NO: 302, SEQ ID NO: 342, SEQ ID NO: 382, SEQ ID NO: 422, SEQ IDNO: 462, SEQ ID NO: 502, SEQ ID NO: 542, SEQ ID NO: 582, SEQ ID NO: 622,SEQ ID NO: 662, SEQ ID NO: 702, SEQ ID NO: 742, SEQ ID NO: 782, SEQ IDNO: 822, SEQ ID NO: 862, SEQ ID NO: 902, SEQ ID NO: 942, or a variantthereof, wherein one or more of the framework residues (FR residues)and/or CDR residues in said V_(H or) V_(L) polypeptide has beensubstituted with another amino acid residue resulting in an anti-PCSK9antibody that specifically binds PCSK9. The invention also includeshumanized and chimeric forms of these antibodies. The chimeric andhumanized antibodies may include an Fc derived from IgG1, IgG2, IgG3,IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, IgG10, IgG11, IgG12, IgG13, IgG14,IgG15, IgG16, IgG17, IgG18 or IgG19 constant regions.

In one embodiment of the invention, the chimeric or humanized antibodiesor fragments or V_(H) or V_(L) polypeptides originate or are derivedfrom one or more rabbit antibodies, e.g., a rabbit antibody isolatedfrom clonal rabbit B cell population.

In some aspects, the invention comprises a vector comprising a nucleicacid molecule encoding an anti-PCSK9 antibody or fragment thereofaccording to the invention. In some embodiments, the invention comprisesa host cell comprising a nucleic acid molecule encoding an anti-PCSK9antibody or fragment thereof according to the invention.

In some aspects, the invention comprises an isolated antibody orantibody fragment according to the invention that competes for bindingto PCSK9 with an antibody or antibody fragment according to theinvention disclosed herein.

In some aspects, the invention comprises a nucleic acid moleculeencoding an antibody or antibody fragment according to the invention.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising at least one antibody or antibody fragmentaccording to the invention.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a patient, comprising administering to a patient in need thereof aneffective amount of at least one isolated antibody or antibody fragmentaccording to the invention.

In some aspects, the invention comprises a method of inhibiting bindingof PCSK9 to LDLR in a subject comprising administering an effectiveamount of at least one antibody or antibody fragment according to theinvention.

In some aspects, the invention comprises an antibody or antibodyfragment according to the invention that selectively binds to PCSK9,wherein the antibody or antibody fragment according to the inventionbinds to PCSK9 with a K_(D) that is less than 100 μM.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, the method comprising administering to a subject in needthereof an effective amount of at least one antibody or antibodyfragment according to the invention simultaneously or sequentially withanother active agent, e.g., an agent that elevates the availability ofLDLR protein or which decreases serum cholesterol.

In some aspects, the invention comprises a method of lowering serumcholesterol level in a subject, the method comprising administering to asubject an effective amount of at least one antibody or antibodyfragment according to the invention as disclosed herein.

In some aspects, the invention relates to the use of the antibodiesdescribed herein, or antibodies competing therewith or possessing thesame or overlapping epitopic specificity, preferably antibodies orantibody fragments comprising one or all of the CDRs of one of theexemplified anti-PCSK9 antibodies or antibody fragments, or morepreferably an antibody comprising one or more variable or CDR sequenceswhich possess at least 80, 90, or 95% identity to any of the VH, VL andCDR polypeptides described herein, and to polynucleotides encoding theseantibodies and antibody fragments and host cells containing. A preferredembodiment of the invention is directed to chimeric or humanizedantibodies and fragments thereof (such as Fab or Fv or monovalentfragments) capable of binding to PCSK9, and preferably which inhibit,block or neutralize the biological activities of PCSK9 or which block orinhibit the binding of PCSK9 to LDLR.

In some aspects, the invention comprises an isolated antibody orantibody fragment that competes for binding to PCSK9 or binds with thesame or an overlapping epitope on PCSK9 as an antibody or antibodyfragment according to the invention.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a patient, comprising administering to a patient in need thereof aneffective amount of at least one anti-PCSK9 antibody or antibodyfragment according to the invention.

In some aspects, the invention comprises a method of inhibiting bindingof PCSK9 to LDLR in a subject in need thereof comprising administeringan effective amount of at least one anti-PCSK9 antibody or antibodyfragment according to the invention.

In some aspects, the invention comprises an anti-PCSK9 antibody orantibody fragment according to the invention that binds to PCSK9 with aKD that is less than 100 nM.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, the method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orantibody fragment according to the invention simultaneously orsequentially with another active agent, e.g., one that reducescholesterol levels or which elevates the availability of LDLR protein.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject, the method comprising administering toa subject an effective amount of at least at least one anti-PCSK9antibody or antibody fragment according to the invention.

In some aspects, the invention comprises a method of lowering serumcholesterol level in a subject, the method comprising administering to asubject an effective amount of at least one anti-PCSK9 antibody orantibody fragment according to the invention, simultaneously orsequentially with another agent that elevates the availability of LDLRprotein.

In some aspects, the invention comprises a method of increasing LDLRprotein level in a subject, the method comprising administering to asubject an effective amount of at least one anti-PCSK9 antibody orantibody fragment according to the invention.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject, the method comprising administering to asubject an effective amount of at least one at least one anti-PCSK9antibody or antibody fragment according to the invention simultaneouslyor sequentially with an agent that elevates the availability of LDLRprotein.

In some aspects, the invention further provides methods of preventing ortreating diseases and disorders associated with PCSK9, e.g., diseasesassociated with increased or decreased levels of PCSK9 and/or mutationsin the PCSK9 gene that affect PCSK9 protein expression, primary sequenceand/or function by administering at least one at least one anti-PCSK9antibody or antibody fragment according to the invention in combinationwith other agents.

In other aspects the present invention provides methods for treating orpreventing disorders of cholesterol or lipid homeostasis and disordersand complications associated therewith, e.g., hypercholesterolemia,hyperlipidemia, hypertriglyceridaemia, sitosterolemia, atherosclerosis,arteriosclerosis, coronary heart disease, metabolic syndrome, acutecoronary syndrome, vascular inflammation, xanthoma and relatedconditions using the subject anti-PCSK9 antibodies and antibodyfragments.

In other specific aspects the present invention provides methods fortreating or preventing disorders of cholesterol or lipid homeostasis anddisorders and complications associated therewith, e.g.,hypercholesterolemia, hyperlipidemia, hypertriglyceridaemia,sitosterolemia, atherosclerosis, arteriosclerosis, coronary heartdisease, metabolic syndrome, acute coronary syndrome, vascularinflammation, xanthoma and other conditions such as obesity,hypertension, diabetes, wherein the subject is treated with the subjectanti-PCSK9 antibodies and antibody fragments, in combination with otherdrugs used to treat such disorders such as e.g., statins, ACEinhibitors, Angiotensin II receptor blockers (ARBs), Antiarrhythmics,Antiplatelet Drugs, aspirin, beta blockers, amiodarone, digoxin,aspirin, anti-clotting agents, digoxin, diuretics, heart failure drugs,vasodilators, blood thinners, other anti-cholesterol drugs such asholestyramine (Questran), gemfibrozil (Lopid, Gemcor), Omacor, andpantethine, other anti-hypertensives, antidiabetigenic drugs such asAlpha-glucosidase inhibitors, Biguanides, Dipeptidyl peptidase-4inhibitors, Insulin therapies, Meglitinides, Sulfonylurea, andThiazolidinediones, and other drugs used to treat conditions wherein thetreated individual may have high cholesterol.

ACE inhibitors may be used in combination with the subject anti-PCSK9antibodies and antibody fragments wherein the moities may be jointly orseparately administered by the same or different means of administrationinclude by way of example: Capoten (captopril), Vasotec (enalapril),Prinivil, Zestril (lisinopril), Lotensin (benazepril), Monopril(fosinopril), Altace (ramipril), Accupril (quinapril), Aceon(perindopril), Mavik (trandolapril), Univasc (moexipril),

ARBs may be used in combination with the subject anti-PCSK9 antibodiesand antibody fragments wherein the moities may be jointly or separatelyadministered by the same or different means of administration include byway of example: Cozaar (losartan), Diovan (valsartan), Avapro(irbesartan), Atacand (candesartan), and Micardis (telmisartan).

Antiarrhythmics may be used in combination with the subject anti-PCSK9antibodies and antibody fragments include by way of example: Tambocor(flecamide), Procanbid (procainamide), Cordarone (amiodarone), andBetapace (sotalol).

Antiplatelet Drugs which may be used in combination with the subjectanti-PCSK9 antibodies and antibody fragments wherein the moities may bejointly or separately administered by the same or different means ofadministration include by way of example:

Anticlotting agents which may be used in combination with the subjectanti-PCSK9 antibodies and antibody fragments wherein the moities may bejointly or separately administered by the same or different means ofadministration include: Tissue plasminogen activator (TPA),Tenecteplase, Alteplase, Urokinase, Reteplase, and Streptokinase.

Beta-blockers may be used in combination with the subject anti-PCSK9antibodies and antibody fragments wherein the moities may be jointly orseparately administered by the same or different means of administrationinclude by way of example: Sectral (acebutolol), Zebeta (bisoprolol),Brevibloc (esmolol), Inderal (propranolol), Tenormin (atenolol),Normodyne, Trandate (labetalol), Coreg (carvedilol), Lopressor, andToprol-XL (metoprolol).

Calcium channel blockers which may be used in combination with thesubject anti-PCSK9 antibodies and antibody fragments wherein the moitiesmay be jointly or separately administered by the same or different meansof administration include by way of example: Norvasc (amlodipine),Plendil (felodipine), Cardizem, Cardizem CD, Cardizem SR, Dilacor XR,Diltia XT, Tiazac (diltiazem), Calan, Calan SR, Covera-HS, Isoptin,Isoptin SR, Verelan, Verelan PM (verapamil), Adalat, Adalat CC,Procardia, Procardia XL (nifedipine), Cardene, Cardene SR (nicardipine),Sular (nisoldipine), Vascor (bepridil), and Caduet which is acombination of a statin cholesterol drug and amlodipine.

Diuretics which may be used in combination with the subject anti-PCSK9antibodies and antibody fragments wherein the moities may be jointly orseparately administered by the same or different means of administrationinclude: by way of example Lasix (furosemide), Bumex (bumetanide),Demadex (torsemide), Esidrix (hydrochlorothiazide), Zaroxolyn(metolazone), and Aldactone (spironolactone).

Heart failure drugs which may be used in combination with the subjectanti-PCSK9 antibodies and antibody fragments wherein the moities may bejointly or separately administered by the same or different means ofadministration include by way of example Dobutrex (dobutamine), andPrimacor (milrinone).

Vasodilators which may be used in combination with the subjectanti-PCSK9 antibodies and antibody fragments wherein the moities may bejointly or separately administered by the same or different means ofadministration include by way of example Dilatrate-SR, Iso-Bid, Isonate,Isorbid, Isordil, Isotrate, Sorbitrate (isosorbide dinitrate), IMDUR(isorbide mononitrate), and BiDil (hydralazine with isosorbidedinitrate.

Blood thinners which may be used in combination with the subjectanti-PCSK9 antibodies and antibody fragments wherein the moities may bejointly or separately administered by the same or different means ofadministration include by way of example Warfarin (coumadin), Heparin,Lovenox, and Fragmin.

In other aspects the present invention further provides methods forimproving blood cholesterol markers associated with increased risk ofheart disease using the subject antibodies and antibody fragments inassociation with any of the foregoing or other actives wherein themoities may be jointly or separately administered by the same ordifferent means of administration. These markers include, but are notlimited to, high total cholesterol, high LDL, high total cholesterol toHDL ratio and high LDL-C to HDL ratio.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody protein or fragment according to the inventionand another active, e.g., one of the actives above-identified, e.g., anagent that elevates the availability of LDLR protein levels or an agentwhich blocks or inhibits cholesterol synthesis. In some embodiments, theagent that blocks or blocks cholesterol synthesis comprises a statin.The statin in some instances potentially may further elevate LDLRlevels. In some embodiments, the statin is selected from the groupconsisting of atorvastatin, cerivastatin, fluvastatin, lovastatin,mevastatin, pitavastatin, pravastatin, rosuvastatin, simvastatin, andsome combination thereof. In some embodiments, the statin is furthercombined with niacin, an absorption inhibitor (ezetimibe), a lipidmodifying agent, or a combination thereof. In some embodiments, theagent that elevates the availability of LDLR protein levels comprises acytokine such as oncostatin M, or a hormone like estrogen, and/or aherbal moiety such as berberine.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody protein or fragment or variant thereof accordingto the invention and an agent that increases high density lipoprotein(HDL) and/or decreases triglyceride levels. In some embodiments, theagent that increases high density lipoprotein (HDL) and/or decreasestriglyceride levels comprises a fibrate. In some embodiments, thefibrate is selected from the group consisting of bezafibrate,ciprofibrate, clofibrate, gemfibrozil, fenofibrate, and some combinationthereof.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody protein or fragment according to the inventionand a bile acid sequestering agent. In some embodiments, the bilesequestering agent is selected from the group consisting ofcholestyramine, colesevelam, colestipol, and some combination thereof.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or antibody fragment according to the inventionand an agent that decreases cholesterol absorption in the intestine. Insome embodiments, the agent that decreases cholesterol absorption in theintestine is ezetimibe.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that decreases cholesterol levels. In some embodiments, the agentthat decreases cholesterol levels is selected from the group consistingof certain anti-psychotic agents, certain HIV protease inhibitors,dietary factors such as high fructose, sucrose, cholesterol or certainfatty acids and certain nuclear receptor agonists and antagonists forRXR, RAR, LXR, FXR.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and a PPARgamma agonist, PPAR alpha/gamma agonist, squalene synthase inhibitor,CETP inhibitor, anti-hypertensive, anti-diabetic agent (such assulphonyl ureas, insulin, GLP-1 analogs, DDPIV inhibitors), ApoBmodulator, MTP inhibitors, arteriosclerosis obliterans treatments, or acombination thereof.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that increases HDL levels. In some embodiments, the agent thatincreases HDL levels is niacin, also known as nicotinic acid. In someembodiments, the niacin is a slow-release formulation.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that inhibits hepatic triglyceride production and/or very lowdensity lipoprotein (VLDL) secretions. In some embodiments, the agentthat inhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions is acipimox.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that decreases lipid absorption in the intestine. In someembodiments, the agent that decreases lipid absorption in the intestineis selected from the group consisting of orlistat, lipstatin, and somecombination thereof.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and anagent that is an anti-hypertensive and/or treats angina. In someembodiments, the agent that is an anti-hypertensive and/or treats anginais amlodipine.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and atleast one other agent, wherein the combination allows for mitigation ofundesirable side-effects of at least one other agent. In someembodiments, the antibody or antibody fragment according to theinvention and the at least one other agent are administeredconcurrently. In some embodiments, the antibody or antibody fragmentaccording to the invention or fragment or variant thereof and at leastone other agent are not administered simultaneously, with the antibodyor antibody fragment according to the invention being administeredbefore or after the at least one other agent is administered.

In some aspects, the invention comprises a pharmaceutical compositioncomprising an antibody or fragment according to the invention and atleast one other agent for treating a condition associated with aberrantcholesterol or for treating a condition wherein the individuals oftenhave high cholesterol. For example an antibody protein or fragment orvariant thereof according to the invention may be combined orco-administered with other drugs such as ACE inhibitors, Angiotensin IIreceptor blockers (ARBs), Antiarrhythmics, Antiplatelet Drugs, aspirin,beta blockers, amiodarone, digoxin, aspirin, anti-clotting agents,digoxin, diuretics, heart failure drugs, vasodilators, blood thinners,other anti-cholesterol drugs such as holestyramine (Questran),gemfibrozil (Lopid, Gemcor), Omacor, and pantethine, otheranti-hypertensives, antidiabetigenic drugs such as Alpha-glucosidaseinhibitors, Biguanides, Dipeptidyl peptidase-4 inhibitors, Insulintherapies, Meglitinides, Sulfonylurea, and Thiazolidinediones, or otherdrugs used to treat conditions wherein the treated individual may havehigh cholesterol. Examples of such drugs are identified supra. In someembodiments, the antibody or fragment according to the invention and atleast one other agent are not administered simultaneously, with theantibody or antibody fragment according to the invention beingadministered before or after the at least one other agent isadministered.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelin a patient comprising administering to a patient in need thereof aneffective amount of at least one isolated antibody or antibody fragmentaccording to the invention as disclosed herein. In some embodiments, thecondition is hypercholesterolemia.

In some aspects, anti-PCSK9 antibodies or fragments according to theinvention bind to PCSK9 with a KD that is less than about 100 nM. Insome embodiments, the anti-PCSK9 antibodies or fragments according tothe invention bind PCSK9 with a KD that is between about 10 and about100 nM. In some embodiments, the anti-PCSK9 antibodies or fragmentsaccording to the invention bind PCSK9 with a KD that is less than about10 nM. In some embodiments, the antibody or fragment that binds PCSK9has a KD that is between about 1 and about 10 nM. In some embodiments,the anti-PCSK9 antibodies or fragments according to the invention thatbinds PCSK9 has a KD that is less than about 1 nM. In some embodiments,the anti-PCSK9 antibodies or fragments according to the invention has aKD that is between about 0.1 and about 1 nM. In some embodiments, theanti-PCSK9 antibodies or fragments according to the invention that bindsPCSK9 has a KD that is between about 0.1 and about 0.5 nM. In someembodiments, the anti-PCSK9 antibodies or fragments according to theinvention binds PCSK9 with a KD that is between about 0.01 and about 0.1nM. In some embodiments, the anti-PCSK9 antibodies or fragmentsaccording to the invention binds PCSK9 with a KD that is between about0.1 and about 10 nM. In some embodiments, the anti-PCSK9 antibodies orfragments according to the invention bind PCSK9 with a KD that isbetween 0.120 and about 7.99 nM.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that decreases cholesterol and which optionally furtherelevates the availability of LDLR protein. In some embodiments, theagent that reduces cholesterol and which optionally elevates theavailability of LDLR protein comprises a statin. In some embodiments,the statin is selected from the group consisting of atorvastatin,cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin,pravastatin, rosuvastatin, simvastatin, and some combination thereof. Insome embodiments, the statin is combined with niacin, an absorptioninhibitor (ezetimibe), a lipid modifying agent, or a combinationthereof. In some embodiments, the agent that elevates the availabilityof LDLR protein levels comprises certain cytokines like oncostatin M,estrogen, and/or certain herbal ingredients such as berberine.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that increases high density lipoprotein (HDL) and/or decreasestriglyceride levels. In some embodiments, the agent that increases highdensity lipoprotein (HDL) and/or decreases triglyceride levels comprisesa fibrate. In some embodiments, the fibrate is selected from the groupconsisting of bezafibrate, ciprofibrate, clofibrate, gemfibrozil,fenofibrate, and some combination thereof.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially witha bile sequestering agent. In some embodiments, the bile sequesteringagent is selected from the group consisting of cholestyramine,colesevelam, colestipol, and some combination thereof.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated anti-PCSK9antibodies or fragments according to the invention simultaneously orsequentially with an agent that decreases cholesterol absorption in theintestine. In some embodiments, the agent that decreases cholesterolabsorption in the intestine is ezetimibe.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that decreases cholesterol levels. In some embodiments, theagent that decreases cholesterol levels is selected from the groupconsisting of certain anti-psychotic agents, certain HIV proteaseinhibitors, dietary factors such as high fructose, sucrose, cholesterolor certain fatty acids and certain nuclear receptor agonists andantagonists for RXR, RAR, LXR, FXR.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that increases HDL levels. In some embodiments, the agent thatincreases HDL levels is niacin, also known as nicotinic acid. In someembodiments, the niacin is a slow-release formulation.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that inhibits hepatic triglyceride production and/or very lowdensity lipoprotein (VLDL) secretions. In some embodiments, the agentthat inhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions is acipimox.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that decreases lipid absorption in the intestine. In someembodiments, the agent that decreases lipid absorption in the intestineis selected from the group consisting of orlistat, lipstatin, and somecombination thereof.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withan agent that is an anti-hypertensive and/or treats angina. In someembodiments, the agent that is an anti-hypertensive and/or treats anginais amlodipine.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention simultaneously or sequentially withat least one other agent, wherein the combination allows for mitigationof undesirable side-effects of at least one other agent. In someembodiments, the anti-PCSK9 antibodies or fragments according to theinvention and the at least one other agent are administeredconcurrently. In some embodiments, the antibody or antibody fragmentaccording to the invention and at least one other agent are notadministered simultaneously, with the anti-PCSK9 antibodies or fragmentsaccording to the invention being administered before or after the atleast one other agent is administered.

In some aspects, the invention comprises a method of lowering the serumcholesterol level in a subject. The method comprises administering to asubject an effective amount of at least one anti-PCSK9 antibody orfragment according to the invention.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat reduces cholesterol and which optionally further optionallyelevates the availability of LDLR protein. In some embodiments, theagent that reduces cholesterol and further optionally elevates theavailability of LDLR protein comprises a statin. In some embodiments,the statin is selected from the group consisting of atorvastatin,cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin,pravastatin, rosuvastatin, simvastatin, and some combination thereof. Insome embodiments, the statin is combined with niacin, an absorptioninhibitor (ezetimibe), a lipid modifying agent, or a combinationthereof. In some embodiments, the agent that elevates the availabilityof LDLR protein levels comprises certain cytokines like oncostatin M,estrogen, and/or certain herbal ingredients such as berberine.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat increases high density lipoprotein (HDL) and/or decreasestriglyceride levels. In some embodiments, the agent that increases highdensity lipoprotein (HDL) and/or decreases triglyceride levels comprisesa fibrate. In some embodiments, the fibrate is selected from the groupconsisting of bezafibrate, ciprofibrate, clofibrate, gemfibrozil,fenofibrate, and some combination thereof.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with a bileacid sequestering agent. In some embodiments, the bile sequesteringagent is selected from the group consisting of cholestyramine,colesevelam, colestipol, and some combination thereof.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one at least one anti-PCSK9 antibody orfragment according to the invention, simultaneously or sequentially withan agent that decreases cholesterol absorption in the intestine. In someembodiments, the agent that decreases cholesterol absorption in theintestine is ezetimibe.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least anti-PCSK9 antibody or fragment accordingto the invention, simultaneously or sequentially with an agent thatdecreases cholesterol levels. In some embodiments, the agent thatdecreases cholesterol levels is selected from the group consisting ofcertain anti-psychotic agents, certain HIV protease inhibitors, dietaryfactors such as high fructose, sucrose, cholesterol or certain fattyacids and certain nuclear receptor agonists and antagonists for RXR,RAR, LXR, FXR.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent thatincreases HDL levels. In some embodiments, the agent that increases HDLlevels is niacin, also known as nicotinic acid. In some embodiments, theniacin is a slow-release formulation.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat inhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions. In some embodiments, the agent thatinhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions is acipimox.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat decreases lipid absorption in the intestine. In some embodiments,the agent that decreases lipid absorption in the intestine is selectedfrom the group consisting of orlistat, lipstatin, and some combinationthereof.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with an agentthat is an anti-hypertensive and/or treats angina. In some embodiments,the agent that is an anti-hypertensive and/or treats angina isamlodipine.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one anti-PCSK9 antibody or fragmentaccording to the invention, simultaneously or sequentially with at leastone other agent, wherein the combination allows for mitigation ofundesirable side-effects of at least one other agent. In someembodiments, the antibody or antibody fragment according to theinvention and the at least one other agent are administeredconcurrently. In some embodiments, the anti-PCSK9 antibody or fragmentaccording to the invention and at least one other agent are notadministered simultaneously, with the antibody or antibody fragmentaccording to the invention being administered before or after the atleast one other agent is administered.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one anti-PCSK9 antibody or fragment according to theinvention as provided herein.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that elevates theavailability of LDLR protein or which reduces cholesterol. In someembodiments, the agent that reduces cholesterol comprises a statin. Insome embodiments, the statin is selected from the group consisting ofatorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin,pitavastatin, pravastatin, rosuvastatin, simvastatin, and somecombination thereof. In some embodiments, the statin is combined withniacin, an absorption inhibitor (ezetimibe), a lipid modifying agent, ora combination thereof. In some embodiments, the agent that elevates theavailability of LDLR protein levels comprises certain cytokines likeoncostatin M, estrogen, and/or certain herbal ingredients such asberberine.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that increaseshigh density lipoprotein (HDL) and/or decreases triglyceride levels. Insome embodiments, the agent that increases high density lipoprotein(HDL) and/or decreases triglyceride levels comprises a fibrate. In someembodiments, the fibrate is selected from the group consisting ofbezafibrate, ciprofibrate, clofibrate, gemfibrozil, fenofibrate, andsome combination thereof.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with a bile acid sequesteringagent. In some embodiments, the bile sequestering agent is selected fromthe group consisting of cholestyramine, colesevelam, colestipol, andsome combination thereof.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that decreasescholesterol absorption in the intestine. In some embodiments, the agentthat decreases cholesterol absorption in the intestine is ezetimibe.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that decreasescholesterol levels. In some embodiments, the agent that decreasescholesterol levels is selected from the group consisting of certainanti-psychotic agents, certain HIV protease inhibitors, dietary factorssuch as high fructose, sucrose, cholesterol or certain fatty acids andcertain nuclear receptor agonists and antagonists for RXR, RAR, LXR,FXR.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that increasesHDL levels. In some embodiments, the agent that increases HDL levels isniacin, also known as nicotinic acid. In some embodiments, the niacin isa slow-release formulation.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that inhibitshepatic triglyceride production and/or very low density lipoprotein(VLDL) secretions. In some embodiments, the agent that inhibits hepatictriglyceride production and/or very low density lipoprotein (VLDL)secretions is acipimox.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that decreaseslipid absorption in the intestine. In some embodiments, the agent thatdecreases lipid absorption in the intestine is selected from the groupconsisting of orlistat, lipstatin, and some combination thereof.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that is ananti-hypertensive and/or treats angina. In some embodiments, the agentthat is an anti-hypertensive and/or treats angina is amlodipine.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with at least one other agent,wherein the combination allows for mitigation of undesirableside-effects of at least one other agent. In some embodiments, theantibody or antibody fragment according to the invention and the atleast one other agent are administered concurrently. In someembodiments, the antibody or antibody fragment according to theinvention and at least one other agent are not administeredsimultaneously, with the antibody or antibody fragment according to theinvention being administered before or after the at least one otheragent is administered.

In some aspects, the invention comprises a method of lowering serumcholesterol level in a subject, the method comprising administering to asubject an effective amount of at least one isolated antibody orantibody fragment according to the invention as disclosed herein,simultaneously or sequentially with another agent that elevates theavailability of LDLR protein or which decreases serum cholesterol.

In some aspects, the invention comprises a method of increasing LDLRprotein level in a subject, the method comprising administering to asubject an effective amount of at least one isolated antibody orantibody fragment according to the invention as disclosed herein.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject, the method comprising administering to asubject an effective amount of at least one isolated antibody orantibody fragment according to the invention as disclosed hereinsimultaneously or sequentially with an agent that elevates theavailability of LDLR protein.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and an agent that blocks or inhibitscholesterol synthesis or which elevates the availability LDLR. In someembodiments, the agent that blocks or inhibits cholesterol synthesiscomprises a statin and may also increase the availability of LDLR. Insome embodiments, the statin is selected from the group consisting ofatorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin,pitavastatin, pravastatin, rosuvastatin, simvastatin, and somecombination thereof. In some embodiments, the statin is combined withniacin, an absorption inhibitor (ezetimibe), a lipid modifying agent, ora combination thereof. In some embodiments, the agent that elevates theavailability of LDLR protein levels comprises certain cytokines likeoncostatin M, estrogen, and/or certain herbal ingredients such asberberine.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and an agent that increases high densitylipoprotein (HDL) and/or decreases triglyceride levels. In someembodiments, the agent that increases high density lipoprotein (HDL)and/or decreases triglyceride levels comprises a fibrate. In someembodiments, the fibrate is selected from the group consisting ofbezafibrate, ciprofibrate, clofibrate, gemfibrozil, fenofibrate, andsome combination thereof.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and a bile acid sequestering agent. Insome embodiments, the bile sequestering agent is selected from the groupconsisting of cholestyramine, colesevelam, colestipol, and somecombination thereof.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and an agent that decreases cholesterolabsorption in the intestine. In some embodiments, the agent thatdecreases cholesterol absorption in the intestine is ezetimibe.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and an agent that decreases cholesterollevels. In some embodiments, the agent that decreases cholesterol levelsis selected from the group consisting of certain anti-psychotic agents,certain HIV protease inhibitors, dietary factors such as high fructose,sucrose, cholesterol or certain fatty acids and certain nuclear receptoragonists and antagonists for RXR, RAR, LXR, FXR.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and an agent that increases HDL levels. Insome embodiments, the agent that increases HDL levels is niacin, alsoknown as nicotinic acid. In some embodiments, the niacin is aslow-release formulation.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and an agent that inhibits hepatictriglyceride production and/or very low density lipoprotein (VLDL)secretions. In some embodiments, the agent that inhibits hepatictriglyceride production and/or very low density lipoprotein (VLDL)secretions is acipimox.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and an agent that decreases lipidabsorption in the intestine. In some embodiments, the agent thatdecreases lipid absorption in the intestine is selected from the groupconsisting of orlistat, lipstatin, and some combination thereof.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and an agent that is an anti-hypertensiveand/or treats angina. In some embodiments, the agent that is ananti-hypertensive and/or treats angina is amlodipine.

In some aspects, the invention comprises a pharmaceutical or diagnosticcomposition comprising an antibody or antibody fragment according to theinvention as disclosed herein and at least one other agent, wherein thecombination allows for mitigation of undesirable side-effects of atleast one other agent. In some embodiments, the antibody or antibodyfragment according to the invention and the at least one other agent areadministered concurrently. In some embodiments, the antibody or antibodyfragment according to the invention and at least one other agent are notadministered simultaneously, with the antibody or antibody fragmentaccording to the invention being administered before or after the atleast one other agent is administered.

In some aspect, the invention comprises a method of making the antibodyor antibody fragment according to the invention as described herein,comprising the step of preparing said antibody or antibody fragmentaccording to the invention from a host cell that secretes said antibodyor antibody fragment according to the invention.

In some aspect, the invention comprises a pharmaceutical or diagnosticcomposition comprising at least one antibody or antibody fragmentaccording to the invention as described herein and a pharmaceuticallyacceptable excipient. In some embodiments, the pharmaceutical ordiagnostic composition further comprises an additional active agent. Insome embodiments, said additional active agent is selected from thegroup consisting of a radioisotope, radionuclide, a toxin, or atherapeutic and a chemotherapeutic group.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with an elevated serum cholesterollevel in a patient. The method comprises administering to a patient inneed thereof an effective amount of at least one isolated antibody orantibody fragment according to the invention as disclosed herein. Insome embodiments, the condition is hypercholesterolemia.

In some aspects, the invention comprises a method of inhibiting bindingof PCSK9 to LDLR in a patient comprising administering an effectiveamount of at least one antibody or antibody fragment according to theinvention according as described herein.

In some aspect, the invention comprises an antibody or antibody fragmentaccording to the invention that binds to PCSK9 with a KD that is lessthan 100 nM. In some embodiments, the antibody or antibody fragmentaccording to the invention binds with a KD that is less than 10 nM. Insome embodiments, the antibody or antibody fragment according to theinvention binds with a KD that is less than 5 nM.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with an agent that elevates theavailability of LDLR protein or which blocks or inhibits cholesterolsynthesis. In some embodiments, the agent that blocks or inhibitscholesterol synthesis comprises a statin. In some embodiments, thestatin is selected from the group consisting of atorvastatin,cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin,pravastatin, rosuvastatin, simvastatin, and some combination thereof. Insome embodiments, the statin is combined with niacin, an absorptioninhibitor (ezetimibe), a lipid modifying agent, or a combinationthereof. In some embodiments, the agent that elevates the availabilityof LDLR protein levels comprises certain cytokines like oncostatin M,estrogen, and/or certain herbal ingredients such as berberine.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with an agent that increases high densitylipoprotein (HDL) and/or decreases triglyceride levels. In someembodiments, the agent that increases high density lipoprotein (HDL)and/or decreases triglyceride levels comprises a fibrate. In someembodiments, the fibrate is selected from the group consisting ofbezafibrate, ciprofibrate, clofibrate, gemfibrozil, fenofibrate, andsome combination thereof.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with a bile acid sequestering agent. Insome embodiments, the bile sequestering agent is selected from the groupconsisting of cholestyramine, colesevelam, colestipol, and somecombination thereof.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with an agent that decreases cholesterolabsorption in the intestine. In some embodiments, the agent thatdecreases cholesterol absorption in the intestine is ezetimibe.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with an agent that decreases cholesterollevels. In some embodiments, the agent that decreases cholesterol levelsis selected from the group consisting of certain anti-psychotic agents,certain HIV protease inhibitors, dietary factors such as high fructose,sucrose, cholesterol or certain fatty acids and certain nuclear receptoragonists and antagonists for RXR, RAR, LXR, FXR.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with an agent that increases HDL levels.In some embodiments, the agent that increases HDL levels is niacin, alsoknown as nicotinic acid. In some embodiments, the niacin is aslow-release formulation.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with an agent that inhibits hepatictriglyceride production and/or very low density lipoprotein (VLDL)secretions. In some embodiments, the agent that inhibits hepatictriglyceride production and/or very low density lipoprotein (VLDL)secretions is acipimox.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with an agent that decreases lipidabsorption in the intestine. In some embodiments, the agent thatdecreases lipid absorption in the intestine is selected from the groupconsisting of orlistat, lipstatin, and some combination thereof.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with an agent that is ananti-hypertensive and/or treats angina. In some embodiments, the agentthat is an anti-hypertensive and/or treats angina is amlodipine.

In some aspects, the invention comprises a method for treating orpreventing a condition associated with elevated serum cholesterol levelsin a subject, said method comprising administering to a subject in needthereof an effective amount of at least one isolated antibody orantibody fragment according to the invention described hereinsimultaneously or sequentially with at least one other agent, whereinthe combination allows for mitigation of undesirable side-effects of atleast one other agent. In some embodiments, the antibody or antibodyfragment according to the invention and the at least one other agent areadministered concurrently. In some embodiments, the antibody or antibodyfragment according to the invention and at least one other agent are notadministered simultaneously, with the antibody or antibody fragmentaccording to the invention being administered before or after the atleast one other agent is administered.

In some aspects, the invention comprises a method of lowering the serumcholesterol level in a subject. The method comprises administering to asubject an effective amount of at least one isolated antibody orantibody fragment according to the invention as described herein.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent thatelevates the availability of LDLR protein or which blocks or inhibitscholesterol synthesis. In some embodiments, the agent that blocks orinhibits cholesterol synthesis comprises a statin. In some embodiments,the statin is selected from the group consisting of atorvastatin,cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin,pravastatin, rosuvastatin, simvastatin, and some combination thereof. Insome embodiments, the statin is combined with niacin, an absorptioninhibitor (ezetimibe), a lipid modifying agent, or a combinationthereof. In some embodiments, the agent that elevates the availabilityof LDLR protein levels comprises certain cytokines like oncostatin M,estrogen, and/or certain herbal ingredients such as berberine.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent thatincreases high density lipoprotein (HDL) and/or decreases triglyceridelevels. In some embodiments, the agent that increases high densitylipoprotein (HDL) and/or decreases triglyceride levels comprises afibrate. In some embodiments, the fibrate is selected from the groupconsisting of bezafibrate, ciprofibrate, clofibrate, gemfibrozil,fenofibrate, and some combination thereof.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with a bile acidsequestering agent. In some embodiments, the bile sequestering agent isselected from the group consisting of cholestyramine, colesevelam,colestipol, and some combination thereof.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent thatdecreases cholesterol absorption in the intestine. In some embodiments,the agent that decreases cholesterol absorption in the intestine isezetimibe.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent thatdecreases cholesterol levels. In some embodiments, the agent thatdecreases cholesterol levels is selected from the group consisting ofcertain anti-psychotic agents, certain HIV protease inhibitors, dietaryfactors such as high fructose, sucrose, cholesterol or certain fattyacids and certain nuclear receptor agonists and antagonists for RXR,RAR, LXR, FXR.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent thatincreases HDL levels. In some embodiments, the agent that increases HDLlevels is niacin, also known as nicotinic acid. In some embodiments, theniacin is a slow-release formulation.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent thatinhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions. In some embodiments, the agent thatinhibits hepatic triglyceride production and/or very low densitylipoprotein (VLDL) secretions is acipimox.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent thatdecreases lipid absorption in the intestine. In some embodiments, theagent that decreases lipid absorption in the intestine is selected fromthe group consisting of orlistat, lipstatin, and some combinationthereof.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with an agent that is ananti-hypertensive and/or treats angina. In some embodiments, the agentthat is an anti-hypertensive and/or treats angina is amlodipine.

In some aspects, the invention comprises a method of lowering serumcholesterol levels in a subject comprising administering to a subject aneffective amount of at least one antibody or antibody fragment accordingto the invention simultaneously or sequentially with at least one otheragent, wherein the combination allows for mitigation of undesirableside-effects of at least one other agent. In some embodiments, theantibody or antibody fragment according to the invention and the atleast one other agent are administered concurrently. In someembodiments, the antibody or antibody fragment according to theinvention and at least one other agent are not administeredsimultaneously, with the antibody or antibody fragment according to theinvention being administered before or after the at least one otheragent is administered.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one isolated antibody or antibody fragment accordingto the invention as provided herein.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that elevates theavailability of LDLR protein or which blocks or inhibits cholesterolsynthesis. In some embodiments, the agent that blocks or inhibitscholesterol synthesis comprises a statin. In some embodiments, thestatin is selected from the group consisting of atorvastatin,cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin,pravastatin, rosuvastatin, simvastatin, and some combination thereof. Insome embodiments, the statin is combined with niacin, an absorptioninhibitor (ezetimibe), a lipid modifying agent, or a combinationthereof. In some embodiments, the agent that elevates the availabilityof LDLR protein levels comprises certain cytokines like oncostatin M,estrogen, and/or certain herbal ingredients such as berberine.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that increaseshigh density lipoprotein (HDL) and/or decreases triglyceride levels. Insome embodiments, the agent that increases high density lipoprotein(HDL) and/or decreases triglyceride levels comprises a fibrate. In someembodiments, the fibrate is selected from the group consisting ofbezafibrate, ciprofibrate, clofibrate, gemfibrozil, fenofibrate, andsome combination thereof.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with a bile acid sequesteringagent. In some embodiments, the bile sequestering agent is selected fromthe group consisting of cholestyramine, colesevelam, colestipol, andsome combination thereof.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that decreasescholesterol absorption in the intestine. In some embodiments, the agentthat decreases cholesterol absorption in the intestine is ezetimibe.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that decreasescholesterol levels. In some embodiments, the agent that decreasescholesterol levels is selected from the group consisting of certainanti-psychotic agents, certain HIV protease inhibitors, dietary factorssuch as high fructose, sucrose, cholesterol or certain fatty acids andcertain nuclear receptor agonists and antagonists for RXR, RAR, LXR,FXR.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that increasesHDL levels. In some embodiments, the agent that increases HDL levels isniacin, also known as nicotinic acid. In some embodiments, the niacin isa slow-release formulation.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that inhibitshepatic triglyceride production and/or very low density lipoprotein(VLDL) secretions. In some embodiments, the agent that inhibits hepatictriglyceride production and/or very low density lipoprotein (VLDL)secretions is acipimox.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that decreaseslipid absorption in the intestine. In some embodiments, the agent thatdecreases lipid absorption in the intestine is selected from the groupconsisting of orlistat, lipstatin, and some combination thereof.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with an agent that is ananti-hypertensive and/or treats angina. In some embodiments, the agentthat is an anti-hypertensive and/or treats angina is amlodipine.

In some aspects, the invention comprises a method of increasing LDLRprotein levels in a subject by administering to a subject an effectiveamount of at least one antibody or antibody fragment according to theinvention simultaneously or sequentially with at least one other agent,wherein the combination allows for mitigation of undesirableside-effects of at least one other agent. In some embodiments, theantibody or antibody fragment according to the invention and the atleast one other agent are administered concurrently. In someembodiments, the antibody or antibody fragment according to theinvention and at least one other agent are not administeredsimultaneously, with the antibody or antibody fragment according to theinvention being administered before or after the at least one otheragent is administered.

In some aspects, the invention comprises a neutralizing antibody thatbinds to PCSK9 and reduces a low density lipoprotein receptor (LDLR)lowering effect of PCSK9 on LDLR. In some embodiments, the antibodyspecifically binds to PCSK9. In some embodiments, the antibody binds tothe catalytic domain of PCSK9.

In some aspects, the invention comprises an isolated neutralizingantibody or antibody fragment according to the invention that binds to aPCSK9 protein comprising the amino acid sequence of SEQ ID NO: 961,wherein the neutralizing antibody or antibody fragment according to theinvention decreases the ability of PCSK9 to lower LDLR levels. In someembodiments, the antibody or antibody fragment according to theinvention is a LDLR non-competitive neutralizing antibody or antibodyfragment according to the invention. In some embodiments, the antibodyor antibody fragment according to the invention is a LDLR competitiveneutralizing antibody or antibody fragment according to the invention.

In some aspects, the invention comprises an isolated neutralizingantibody or antibody fragment according to the invention that binds to aPCSK9 protein comprising the amino acid sequence of SEQ ID NO: 962,wherein the neutralizing antibody or antibody fragment according to theinvention decreases the LDLR lowering effect of PCSK9 on LDLR. In someembodiments, the antibody or antibody fragment according to theinvention is a LDLR non-competitive neutralizing antibody or antibodyfragment according to the invention. In some embodiments, the antibodyor antibody fragment according to the invention is a LDLR competitiveneutralizing antibody or antibody fragment according to the invention.

In some aspects, the invention comprises a composition comprising acrystallized PCSK9 protein and an antibody or antibody fragmentaccording to the invention that binds to PCSK9. The compositioncomprises the crystallized PCSK9 protein is such that the threedimensional structure of the PCSK9 protein can be determined to aresolution of about 2.2 angstroms or better. In some embodiments, theantibody or antibody fragment according to the invention is an antibodyor a fragment thereof.

In some aspects, the invention comprises the use of an antibody orantibody fragment according to the invention as described herein, in thepreparation of a medicament for the lowering of serum cholesterol.

In some aspects, the invention comprises the use of an antibody orantibody fragment according to the invention as described herein, in thepreparation of a medicament for treating or preventing a conditionassociated with elevated serum cholesterol levels in a subject.

The inventive antibodies and fragments thereof may be modifiedpost-translationally to add effector moieties such as chemical linkers,detectable moieties such as for example fluorescent dyes, enzymes,substrates, bioluminescent materials, radioactive materials, andchemiluminescent moieties, or functional moieties such as for examplestreptavidin, avidin, biotin, a cytotoxin, a cytotoxic agent, andradioactive materials.

Antibodies or fragments thereof may also be chemically modified toprovide additional advantages such as increased solubility, stabilityand circulating time (in vivo half-life) of the polypeptide, ordecreased immunogenicity (See U.S. Pat. No. 4,179,337). The chemicalmoieties for derivitization may be selected from water soluble polymerssuch as polyethylene glycol, ethylene glycol/propylene glycolcopolymers, carboxymethylcellulose, dextran, polyvinyl alcohol and thelike. The antibodies and fragments thereof may be modified at randompositions within the molecule, or at predetermined positions within themolecule and may include one, two, three or more attached chemicalmoieties.

The polymer may be of any molecular weight, and may be branched orunbranched. For polyethylene glycol, the preferred molecular weight isbetween about 1 kDa and about 100 kDa (the term “about” indicating thatin preparations of polyethylene glycol, some molecules will weigh more,some less, than the stated molecular weight) for ease in handling andmanufacturing. Other sizes may be used, depending on the desiredtherapeutic profile (e.g., the duration of sustained release desired,the effects, if any on biological activity, the ease in handling, thedegree or lack of antigenicity and other known effects of thepolyethylene glycol to a therapeutic protein or analog). For example,the polyethylene glycol may have an average molecular weight of about200, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500,6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000,11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500,16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, 20,000,25,000, 30,000, 35,000, 40,000, 50,000, 55,000, 60,000, 65,000, 70,000,75,000, 80,000, 85,000, 90,000, 95,000, or 100,000 kDa. Branchedpolyethylene glycols are described, for example, in U.S. Pat. No.5,643,575; Morpurgo et al., Appl. Biochem. Biotechnol. 56:59-72 (1996);Vorobjev et al., Nucleosides Nucleotides 18:2745-2750 (1999); andCaliceti et al., Bioconjug. Chem. 10:638-646 (1999), the disclosures ofeach of which are incorporated herein by reference.

There are a number of attachment methods available to those skilled inthe art, See e.g., EP 0 401 384, herein incorporated by reference(coupling PEG to G-CSF), See also Malik et al., Exp. Hematol.20:1028-1035 (1992) (reporting pegylation of GM-CSF using tresylchloride). For example, polyethylene glycol may be covalently boundthrough amino acid residues via a reactive group, such as, a free aminoor carboxyl group. Reactive groups are those to which an activatedpolyethylene glycol molecule may be bound. The amino acid residueshaving a free amino group may include lysine residues and the N-terminalamino acid residues; those having a free carboxyl group may includeaspartic acid residues glutamic acid residues and the C-terminal aminoacid residue. Sulfhydryl groups may also be used as a reactive group forattaching the polyethylene glycol molecules. Preferred for therapeuticpurposes is attachment at an amino group, such as attachment at theN-terminus or lysine group.

As suggested above, polyethylene glycol may be attached to proteins vialinkage to any of a number of amino acid residues. For example,polyethylene glycol can be linked to polypeptides via covalent bonds tolysine, histidine, aspartic acid, glutamic acid, or cysteine residues.One or more reaction chemistries may be employed to attach polyethyleneglycol to specific amino acid residues (e.g., lysine, histidine,aspartic acid, glutamic acid, or cysteine) or to more than one type ofamino acid residue (e.g., lysine, histidine, aspartic acid, glutamicacid, cysteine and combinations thereof).

Alternatively, antibodies or fragments thereof may have increased invivo half lives via fusion with albumin (including but not limited torecombinant human serum albumin or fragments or variants thereof (See,e.g., U.S. Pat. No. 5,876,969, issued Mar. 2, 1999, EP Patent 0 413 622,and U.S. Pat. No. 5,766,883, issued Jun. 16, 1998, herein incorporatedby reference in their entirety)) or other circulating blood proteinssuch as transferrin or ferritin. In a preferred embodiment, polypeptidesand/or antibodies of the present invention (including fragments orvariants thereof) are fused with the mature form of human serum albumin(i.e., amino acids 1-585 of human serum albumin as shown in FIGS. 1 and2 of EP Patent 0 322 094) which is herein incorporated by reference inits entirety. Polynucleotides encoding fusion proteins of the inventionare also encompassed by the invention.

Regarding detectable moieties, further exemplary enzymes include, butare not limited to, horseradish peroxidase, acetylcholinesterase,alkaline phosphatase, beta-galactosidase and luciferase. Furtherexemplary fluorescent materials include, but are not limited to,rhodamine, fluorescein, fluorescein isothiocyanate, umbelliferone,dichlorotriazinylamine, phycoerythrin, and dansyl chloride. Furtherexemplary chemiluminescent moieties include, but are not limited to,luminol. Further exemplary bioluminescent materials include, but are notlimited to, luciferin and aequorin. Further exemplary radioactivematerials include, but are not limited to, Iodine 125 (¹²⁵I), Carbon 14(¹⁴C), Sulfur 35 (³⁵S), Tritium (³H) and Phosphorus 32 (³²P).

Regarding functional moieties, exemplary cytotoxic agents include, butare not limited to, methotrexate, aminopterin, 6-mercaptopurine,6-thioguanine, cytarabine, 5-fluorouracil decarbazine; alkylating agentssuch as mechlorethamine, thioepa chlorambucil, melphalan, carmustine(BSNU), mitomycin C, lomustine (CCNU), 1-methylnitrosourea,cyclothosphamide, mechlorethamine, busulfan, dibromomannitol,streptozotocin, mitomycin C, cis-dichlorodiamine platinum (II) (DDP)cisplatin and carboplatin (paraplatin); anthracyclines includedaunorubicin (formerly daunomycin), doxorubicin (adriamycin),detorubicin, caminomycin, idarubicin, epirubicin, mitoxantrone andbisantrene; antibiotics include dactinomycin (actinomycin D), bleomycin,calicheamicin, mithramycin, and anthramycin (AMC); and antimytoticagents such as the vinca alkaloids, vincristine and vinblastine. Othercytotoxic agents include paclitaxel (taxol), ricin, pseudomonasexotoxin, gemcitabine, cytochalasin B, gramicidin D, ethidium bromide,emetine, etoposide, tenoposide, colchicin, dihydroxy anthracin dione,1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine,propranolol, puromycin, procarbazine, hydroxyurea, asparaginase,corticosteroids, mytotane (O,P′-(DDD)), interferons, and mixtures ofthese cytotoxic agents.

Further cytotoxic agents include, but are not limited to,chemotherapeutic agents such as carboplatin, cisplatin, paclitaxel,gemcitabine, calicheamicin, doxorubicin, 5-fluorouracil, mitomycin C,actinomycin D, cyclophosphamide, vincristine and bleomycin. Toxicenzymes from plants and bacteria such as ricin, diphtheria toxin andPseudomonas toxin may be conjugated to the humanized or chimericantibodies, or binding fragments thereof, to generatecell-type-specific-killing reagents (Youle, et al., Proc. Nat'l Acad.Sci. USA 77:5483 (1980); Gilliland, et al., Proc. Nat'l Acad. Sci. USA77:4539 (1980); Krolick, et al., Proc. Nat'l Acad. Sci. USA 77:5419(1980)).

Other cytotoxic agents include cytotoxic ribonucleases as described byGoldenberg in U.S. Pat. No. 6,653,104. Embodiments of the invention alsorelate to radioimmunoconjugates where a radionuclide that emits alpha orbeta particles is stably coupled to the antibody, or binding fragmentsthereof, with or without the use of a complex-forming agent. Suchradionuclides include beta-emitters such as Phosphorus-32 (³²P),Scandium-47 (⁴⁷Sc), Copper-67 (⁶⁷Cu), Gallium-67 (⁶⁷Ga), Yttrium-88(⁸⁸Y), Yttrium-90 (⁹⁰Y), Iodine-125 (¹²⁵I), Iodine-131 (¹³¹I),Samarium-153 (¹⁵³Sm), Lutetium-177 (¹⁷⁷Lu), Rhenium-186 (¹⁸⁶Re) orRhenium-188 (¹⁸⁸Re), and alpha-emitters such as Astatine-211 (²¹¹At),Lead-212 (²¹²Pb), Bismuth-212 (²¹²Bi) or -213 (²¹³Bi) or Actinium-225(²²⁵Ac).

Methods are known in the art for conjugating an antibody or bindingfragment thereof to a detectable moiety and the like, such as forexample those methods described by Hunter et al, Nature 144:945 (1962);David et al, Biochemistry 13:1014 (1974); Pain et al, J. Immunol. Meth.40:219 (1981); and Nygren, J., Histochem. and Cytochem. 30:407 (1982).

Embodiments described herein further include variants and equivalentsthat are substantially homologous to the antibodies, antibody fragments,diabodies, SMIPs, camelbodies, nanobodies, IgNAR, polypeptides, variableregions and CDRs set forth herein. These may contain, e.g., conservativesubstitution mutations, (i.e., the substitution of one or more aminoacids by similar amino acids). For example, conservative substitutionrefers to the substitution of an amino acid with another within the samegeneral class, e.g., one acidic amino acid with another acidic aminoacid, one basic amino acid with another basic amino acid, or one neutralamino acid by another neutral amino acid. What is intended by aconservative amino acid substitution is well known in the art.

In another embodiment, the invention contemplates polypeptide sequenceshaving at least 90% or greater sequence homology to any one or more ofthe polypeptide sequences of antibody fragments, variable regions andCDRs set forth herein. More preferably, the invention contemplatespolypeptide sequences having at least 95% or greater sequence homology,even more preferably at least 98% or greater sequence homology, andstill more preferably at least 99% or greater sequence homology to anyone or more of the polypeptide sequences of antibody fragments, variableregions and CDRs set forth herein. Methods for determining homologybetween nucleic acid and amino acid sequences are well known to those ofordinary skill in the art.

In another embodiment, the invention further contemplates theabove-recited polypeptide homologs of the antibody fragments, variableregions and CDRs set forth herein further having anti-PCSK9 activity.Non-limiting examples of anti-PCSK9 activity are set forth herein.

In another embodiment, the invention further contemplates the generationand use of anti-idiotypic antibodies that bind any of the foregoingsequences. In an exemplary embodiment, such an anti-idiotypic antibodycould be administered to a subject who has received an anti-PCSK9antibody to modulate, reduce, or neutralize, the effect of theanti-PCSK9 antibody. Such anti-idiotypic antibodies could also be usefulfor treatment of an autoimmune disease characterized by the presence ofanti-PCSK9 antibodies. A further exemplary use of such anti-idiotypicantibodies is for detection of the anti-PCSK9 antibodies of the presentinvention, for example to monitor the levels of the anti-PCSK9antibodies present in a subject's blood or other bodily fluids.

The present invention also contemplates anti-PCSK9 antibodies comprisingany of the polypeptide or polynucleotide sequences described hereinsubstituted for any of the other polynucleotide sequences describedherein. For example, without limitation thereto, the present inventioncontemplates antibodies comprising the combination of any of thevariable light chain and variable heavy chain sequences describedherein, and further contemplates antibodies resulting from substitutionof any of the CDR sequences described herein for any of the other CDRsequences described herein.

Additional Exemplary Embodiments of the Invention

In another embodiment, the invention contemplates one or more anti-humanPCSK9 antibodies or antibody fragments thereof which specifically bindto the same linear or conformational epitope(s) and/or competes forbinding to the same or overlapping linear or conformational epitope(s)on an intact human PCSK9 polypeptide or fragment thereof as ananti-human PCSK9 antibody selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18,Ab19, Ab20, Ab21, Ab22, Ab23, or Ab24. In a preferred embodiment, theanti-human PCSK9 antibody or fragment thereof specifically binds to thesame linear or conformational epitope(s) and/or competes for binding tothe same linear or conformational epitope(s) on an intact human PCSK9polypeptide or a fragment thereof as Ab7.

A preferred embodiment of the invention is directed to chimeric orhumanized antibodies and fragments thereof (including Fab fragments)having binding specificity for PCSK9 and inhibiting biologicalactivities mediated by the binding of PCSK9 to the LDLR.

In certain instances, PCSK9 activity correlates with a number of humandisease states. For example, in certain instances, too much or toolittle PCSK9 activity correlates with certain conditions, such ashypercholesterolemia. Therefore, in certain instances, modulating PCSK9activity can be therapeutically useful. In certain embodiments, aneutralizing antibody or antibody fragment according to the invention toPCSK9 is used to modulate at least one PCSK9 activity (e.g., binding toLDLR). Such methods can treat and/or prevent and/or reduce the risk ofdisorders that relate to elevated serum cholesterol levels or in whichelevated cholesterol levels are relevant.

As will be appreciated by one of skill in the art, in light of thepresent disclosure, disorders that relate to, involve, or can beinfluenced by varied cholesterol, LDL, or LDLR levels can be addressedby various embodiments of the antibody or antibody fragment according tothe invention. In some embodiments, a “cholesterol related disorder”(which includes “serum cholesterol related disorders”) includes any oneor more of the following: hypercholesterolemia, heart disease, metabolicsyndrome, diabetes, coronary heart disease, stroke, cardiovasculardiseases, Alzheimer's disease and generally dyslipidemias, which can bemanifested, for example, by an elevated total serum cholesterol,elevated LDL, elevated triglycerides, elevated VLDL, and/or low HDL.Some non-limiting examples of primary and secondary dyslipidemias thatcan be treated using an antibody or antibody fragment according to theinvention, either alone, or in combination with one or more other agentsinclude the metabolic syndrome, diabetes mellitus, familial combinedhyperlipidemia, familial hypertriglyceridemia, familialhypercholesterolemias, including heterozygous hypercholesterolemia,homozygous hypercholesterolemia, familial defective apoplipoproteinB-100; polygenic hypercholesterolemia; remnant removal disease, hepaticlipase deficiency; dyslipidemia secondary to any of the following:dietary indiscretion, hypothyroidism, drugs including estrogen andprogestin therapy, beta-blockers, and thiazide diuretics; nephroticsyndrome, chronic renal failure, Cushing's syndrome, primary biliarycirrhosis, glycogen storage diseases, hepatoma, cholestasis, acromegaly,insulinoma, isolated growth hormone deficiency, and alcohol-inducedhypertriglyceridemia. antibody or antibody fragment according to theinvention can also be useful in preventing or treating atheroscleroticdiseases, such as, for example, coronary heart disease, coronary arterydisease, peripheral arterial disease, stroke (ischaemic andhemorrhagic), angina pectoris, or cerebrovascular disease and acutecoronary syndrome, myocardial infarction. In some embodiments, theantibody or antibody fragment according to the invention is useful inreducing the risk of: nonfatal heart attacks, fatal and non-fatalstrokes, certain types of heart surgery, hospitalization for heartfailure, chest pain in patients with heart disease, and/orcardiovascular events because of established heart disease such as priorheart attack, prior heart surgery, and/or chest pain with evidence ofclogged arteries. In some embodiments, the antibody or antibody fragmentaccording to the invention and methods can be used to reduce the risk ofrecurrent cardiovascular events.

As will be appreciated by one of skill in the art, diseases or disordersthat are generally addressable (either treatable or preventable) throughthe use of statins can also benefit from the application of the instantantibody or antibody fragment according to the invention. In addition,in some embodiments, disorders or disease that can benefit from theprevention of cholesterol synthesis or increased LDLR expression canalso be treated by various embodiments of the antibody or antibodyfragment according to the invention. In addition, as will be appreciatedby one of skill in the art, the use of the anti-PCSK9 antibodies can beespecially useful in the treatment of diabetes. Not only is diabetes arisk factor for coronary heart disease, but insulin increases theexpression of PCSK9. That is, people with diabetes have elevated plasmalipid levels (which can be related to high PCSK9 levels) and can benefitfrom lowering those levels. This is generally discussed in more detailin Costet et al. (“Hepatic PCSK9 Expression is Regulated by NutritionalStatus via Insulin and Sterol Regulatory Element-binding Protein 1C”, J.Biol. Chem., 281: 6211-6218, 2006), the entirety of which isincorporated herein by reference.

In some embodiments, the antibody or antibody fragment according to theinvention is administered to those who have diabetes mellitus, abdominalaortic aneurysm, atherosclerosis and/or peripheral vascular disease inorder to decrease their serum cholesterol levels to a safer range. Insome embodiments, the antibody or antibody fragment according to theinvention is administered to patients at risk of developing any of theherein described disorders. In some embodiments, the antibody orantibody fragment according to the invention are administered tosubjects that smoke, have hypertension or a familial history of earlyheart attacks.

In some embodiments, a subject is administered an antibody or antibodyfragment according to the invention if they are at a moderate risk orhigher on the 2004 NCEP treatment goals. In some embodiments, theantibody or antibody fragment according to the invention is administeredto a subject if the subject's LDL-C cholesterol level is greater than160 mg/dL. In some embodiments, the antibody or antibody fragmentaccording to the invention is administered if the subjects LDL-Ccholesterol level is greater than 130 (and they have a moderate ormoderately high risk according to the 2004 NCEP treatment goals). Insome embodiments, the antibody or antibody fragment according to theinvention is administered if the subjects LDL-C cholesterol level isgreater than 100 (and they have a high or very high risk according tothe 2004 NCEP treatment goals).

A physician will be able to select an appropriate treatment indicationsand target lipid levels depending on the individual profile of aparticular patient. One well-accepted standard for guiding treatment ofhyperlipidemia is the Third Report of the National Cholesterol EducationProgram (NCEP) Expert Panel on Detection, Evaluation, and Treatment ofthe High Blood Cholesterol in Adults (Adult Treatment Panel III) FinalReport, National Institutes of Health, NIH Publication No. 02-5215(2002), the printed publication of which is hereby incorporated byreference in its entirety.

In some embodiments, antibody or antibody fragment according to theinvention to PCSK9 are used to decrease the amount of PCSK9 activityfrom an abnormally high level or even a normal level. In someembodiments, antibody or antibody fragment according to the invention toPCSK9 are used to treat or prevent hypercholesterolemia and/or in thepreparation of medicaments therefore and/or for other cholesterolrelated disorders (such as those noted herein). In certain embodiments,an antibody or antibody fragment according to the invention to PCSK9 isused to treat or prevent conditions such as hypercholesterolemia inwhich PCSK9 activity is normal. In such conditions, for example,reduction of PCSK9 activity to below normal can provide a therapeuticeffect.

In some embodiments, more than one antibody or antibody fragmentaccording to the invention to PCSK9 is used to modulate PCSK9 activity.

In certain embodiments, methods are provided of treating a cholesterolrelated disorder, such as hypercholesterolemia comprising administeringa therapeutically effective amount of one or more antibody or antibodyfragment according to the invention to PCSK9 and another therapeuticagent.

Pharmaceutical or diagnostic compositions of the invention can beadministered in combination therapy, i.e., combined with other agents.In certain embodiments, the combination therapy comprises an antibody orantibody fragment according to the invention capable of binding PCSK9,in combination with at least one anti-cholesterol agent. Agents include,but are not limited to, in vitro synthetically prepared chemicalcompositions, antibodies, antigen binding regions, and combinations andconjugates thereof. In certain embodiments, an agent can act as anagonist, antagonist, allosteric modulator, or toxin. In certainembodiments, an agent can act to inhibit or stimulate its target (e.g.,receptor or enzyme activation or inhibition), and thereby promoteincreased expression of LDLR or decrease serum cholesterol levels.

In certain embodiments, an antibody or antibody fragment according tothe invention to PCSK9 can be administered prior to, concurrent with,and subsequent to treatment with a cholesterol-lowering (serum and/ortotal cholesterol) agent. In certain embodiments, an antibody orantibody fragment according to the invention to PCSK9 can beadministered prophylactically to prevent or mitigate the onset ofhypercholesterolemia, heart disease, diabetes, and/or any of thecholesterol related disorder. In certain embodiments, an antibody orantibody fragment according to the invention to PCSK9 can beadministered for the treatment of an existing hypercholesterolemiacondition. In some embodiments, the antibody or antibody fragmentaccording to the invention delays the onset of the disorder and/orsymptoms associated with the disorder. In some embodiments, the antibodyor antibody fragment according to the invention is provided to a subjectlacking any symptoms of any one of the cholesterol related disorders ora subset thereof.

In certain embodiments, an antibody or antibody fragment according tothe invention to PCSK9 is used with particular therapeutic agents totreat various cholesterol related disorders, such ashypercholesterolemia. In certain embodiments, in view of the conditionand the desired level of treatment, two, three, or more agents can beadministered. In certain embodiments, such agents can be providedtogether by inclusion in the same formulation. In certain embodiments,such agent(s) and an antibody or antibody fragment according to theinvention to PCSK9 can be provided together by inclusion in the sameformulation. In certain embodiments, such agents can be formulatedseparately and provided together by inclusion in a treatment kit. Incertain embodiments, such agents and an antibody or antibody fragmentaccording to the invention to PCSK9 can be formulated separately andprovided together by inclusion in a treatment kit. In certainembodiments, such agents can be provided separately. In certainembodiments, when administered by gene therapy, the genes encodingprotein agents and/or an antibody or antibody fragment according to theinvention to PCSK9 can be included in the same vector. In certainembodiments, the genes encoding protein agents and/or an antibody orantibody fragment according to the invention to PCSK9 can be under thecontrol of the same promoter region. In certain embodiments, the genesencoding protein agents and/or an antibody or antibody fragmentaccording to the invention to PCSK9 can be in separate vectors.

In certain embodiments, the invention provides for pharmaceutical ordiagnostic compositions comprising an antibody or antibody fragmentaccording to the invention to PCSK9 together with a pharmaceuticallyacceptable diluent, carrier, solubilizer, emulsifier, or preservative.

In certain embodiments, the invention provides for pharmaceutical ordiagnostic compositions comprising an antibody or antibody fragmentaccording to the invention to PCSK9 and a therapeutically effectiveamount of at least one additional therapeutic agent, together with apharmaceutically acceptable diluent, carrier, solubilizer, emulsifier,preservative and/or adjuvant.

In certain embodiments, an antibody or antibody fragment according tothe invention to PCSK9 can be used with at least one therapeutic agentfor inflammation. In certain embodiments, an antibody or antibodyfragment according to the invention to PCSK9 can be used with at leastone therapeutic agent for an immune disorder. Exemplary therapeuticagents for inflammation and immune disorders include, but are notlimited to cyclooxygenase type 1 (COX-1) and cyclooxygenase type 2(COX-2) inhibitors, small molecule modulators of 38 kDamitogen-activated protein kinase (p38-MAPK); small molecule modulatorsof intracellular molecules involved in inflammation pathways, whereinsuch intracellular molecules include, but are not limited to, jnk, IKK,NF-κB, ZAP70, and lck. Certain exemplary therapeutic agents forinflammation are described, e.g., in C. A. Dinarello & L. L. MoldawerProinflammatory and Anti-Inflammatory Cytokines in Rheumatoid Arthritis:A Primer for Clinicians Third Edition (2001) Amgen Inc. Thousand Oaks,Calif.

In certain embodiments, pharmaceutical or diagnostic compositions willinclude more than one different antibody or antibody fragment accordingto the invention to PCSK9. In certain embodiments, pharmaceutical ordiagnostic compositions will include more than one antibody or antibodyfragment according to the invention to PCSK9 wherein the antibody orantibody fragment according to the invention to PCSK9 bind more than oneepitope. In some embodiments, the various antibody or antibody fragmentaccording to the invention will not compete with one another for bindingto PCSK9.

In certain embodiments, acceptable formulation materials preferably arenontoxic to recipients at the dosages and concentrations employed. Insome embodiments, the formulation material(s) are for s.c. and/or I.V.administration. In certain embodiments, the pharmaceutical or diagnosticcomposition can contain formulation materials for modifying, maintainingor preserving, for example, the pH, osmolarity, viscosity, clarity,color, isotonicity, odor, sterility, stability, rate of dissolution orrelease, adsorption or penetration of the composition. In certainembodiments, suitable formulation materials include, but are not limitedto, amino acids (such as glycine, glutamine, asparagine, arginine orlysine); antimicrobials; antioxidants (such as ascorbic acid, sodiumsulfite or sodium hydrogen-sulfite); buffers (such as borate,bicarbonate, Tris-HCl, citrates, phosphates or other organic acids);bulking agents (such as mannitol or glycine); chelating agents (such asethylenediamine tetraacetic acid (EDTA)); complexing agents (such ascaffeine, polyvinylpyrrolidone, beta-cyclodextrin orhydroxypropyl-beta-cyclodextrin); fillers; monosaccharides;disaccharides; and other carbohydrates (such as glucose, mannose ordextrins); proteins (such as serum albumin, gelatin or immunoglobulins);coloring, flavoring and diluting agents; emulsifying agents; hydrophilicpolymers (such as polyvinylpyrrolidone); low molecular weightpolypeptides; salt-forming counterions (such as sodium); preservatives(such as benzalkonium chloride, benzoic acid, salicylic acid,thimerosal, phenethyl alcohol, methylparaben, propylparaben,chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such asglycerin, propylene glycol or polyethylene glycol); sugar alcohols (suchas mannitol or sorbitol); suspending agents; surfactants or wettingagents (such as pluronics, PEG, sorbitan esters, polysorbates such aspolysorbate 20, polysorbate 80, triton, tromethamine, lecithin,cholesterol, tyloxapal); stability enhancing agents (such as sucrose orsorbitol); tonicity enhancing agents (such as alkali metal halides,preferably sodium or potassium chloride, mannitol sorbitol); deliveryvehicles; diluents; excipients and/or pharmaceutical adjuvants.(Remington's Pharmaceutical Sciences, 18 th Edition, A. R. Gennaro, ed.,Mack Publishing Company (1995). In some embodiments, the formulationcomprises PBS; 20 mM NaOAC, pH 5.2, 50 mM NaCl; and/or 10 mM NAOAC, pH5.2, 9% Sucrose.

The invention is also directed to an anti-PCSK9 antibody that binds withthe same PCSK9 epitope and/or competes with an anti-PCSK9 antibody forbinding to PCSK9 as an antibody or antibody fragment disclosed herein,including but not limited to an anti-PCSK9 antibody selected from Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14,Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, or Ab24.

In another embodiment, the invention is also directed to an isolatedanti-PCSK9 antibody or antibody fragment comprising one or more of theCDRs contained in the V_(H) polypeptide sequences selected from: SEQ IDNO: 2, SEQ ID NO: 42, SEQ ID NO: 82, SEQ ID NO: 122, SEQ ID NO: 162, SEQID NO: 202, SEQ ID NO: 242, SEQ ID NO: 282, SEQ ID NO: 322, SEQ ID NO:362, SEQ ID NO: 402, SEQ ID NO: 442, SEQ ID NO: 482, SEQ ID NO: 522, SEQID NO: 562, SEQ ID NO: 602, SEQ ID NO: 642, SEQ ID NO: 682, SEQ ID NO:722, SEQ ID NO: 762, SEQ ID NO: 802, SEQ ID NO: 842, SEQ ID NO: 882, SEQID NO: 922, or a variant thereof, and/or one or more of the CDRscontained in the V_(L) polypeptide sequences selected from: SEQ ID NO:22, SEQ ID NO: 62, SEQ ID NO: 102, SEQ ID NO: 142, SEQ ID NO: 182, SEQID NO: 222, SEQ ID NO: 262, SEQ ID NO: 302, SEQ ID NO: 342, SEQ ID NO:382, SEQ ID NO: 422, SEQ ID NO: 462, SEQ ID NO: 502, SEQ ID NO: 542, SEQID NO: 582, SEQ ID NO: 622, SEQ ID NO: 662, SEQ ID NO: 702, SEQ ID NO:742, SEQ ID NO: 782, SEQ ID NO: 822, SEQ ID NO: 862, SEQ ID NO: 902, SEQID NO: 942, or a variant thereof.

In one embodiment of the invention, the anti-human PCSK9 antibodycomprises at least 2 complementarity determining regions (CDRs) in eachthe variable light and the variable heavy regions which are identical tothose contained in an anti-human PCSK9 antibody selected from Ab1, Ab2,Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15,Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, or Ab24.

In a preferred embodiment, the anti-human PCSK9 antibody comprises atleast 2 complementarity determining regions (CDRs) in each the variablelight and the variable heavy regions which are identical to the CDRscontained in an anti-human PCSK9 antibody selected from Ab1, Ab2, Ab3,Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16,Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, or Ab24. In anotherembodiment, all of the CDRs of the anti-human PCSK9 antibody areidentical to the CDRs contained in an anti-human PCSK9 antibody selectedfrom Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12,Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, orAb24.

The invention further contemplates that the one or more anti-human PCSK9antibodies discussed above are aglycosylated; or lack or substantiallylack N and/r O-glyosylation, e.g., as the result of an Fc regioncodification. Also, the subject antibodies may contain an Fc region thathas been modified to alter the effector function, half-life, proteolysisor other properties of the antibody; especially human, humanized orchimeric antibodies according to the invention may contain such modifiedFc regions.

The invention further contemplates one or more anti-human PCSK9antibodies wherein the framework regions (FRs) in the variable lightregion and the variable heavy regions of said antibody respectively arehuman FRs which are unmodified or which have been modified by thesubstitution of one or more human FR residues in the variable light orheavy chain region with the corresponding FR residues of the parentrabbit antibody, and wherein said human FRs have been derived from humanvariable heavy and light chain antibody sequences which have beenselected from a library of human germline antibody sequences based ontheir high level of homology to the corresponding rabbit variable heavyor light chain regions relative to other human germline antibodysequences contained in the library.

In one embodiment of the invention, the anti-human PCSK9 antibody orfragment specifically binds to PCSK9 expressing human cells and/or tocirculating soluble PCSK9 molecules in vivo, including PCSK9 expressedon or by human cells in a patient with a disease associated with cellsthat express PCSK9.

The invention further contemplates anti-human PCSK9 antibodies orfragments directly or indirectly attached to a detectable label ortherapeutic agent.

The invention also contemplates one or more nucleic acid sequences whichresult in the expression of an anti-human PCSK9 antibody or antibodyfragment as set forth above, including those comprising, oralternatively consisting of, yeast or human preferred codons. Theinvention also contemplates vectors (including plasmids or recombinantviral vectors) comprising said nucleic acid sequence(s). The inventionalso contemplates host cells or recombinant host cells expressing atleast one of the antibodies set forth above, including e.g., mammalian,yeast, fungal, bacterial, plant, avian, and insect cells. In a preferredembodiment, the host cell is a yeast or fungal or mammalian cell. In afurther preferred embodiment, the yeast cell is a diploidal yeast cell.In a more preferred embodiment, the yeast cell is a Pichia yeast.

The anti-PCSK9 activity of the anti-PCSK9 antibodies of the presentinvention, and fragments thereof having binding specificity to PCSK9,may also be described by their strength of binding or their affinity forPCSK9. In one embodiment of the invention, the anti-PCSK9 antibodies ofthe present invention, and fragments thereof having binding specificityto PCSK9, bind to PCSK9 with a dissociation constant (K_(d)) of lessthan or equal to 10⁻² M, 5×10⁻³ M, 10⁻³ M, 5×10⁻⁴ M, 10⁻⁴ M, 5×10⁻⁵ M,10⁻⁵ M, 5×10⁻⁶ M, 10⁻⁶ M, 5×10⁻⁷ M, 10⁻⁷ M, 5×10⁻⁸ M, 10⁻⁸ M, 5×10⁻⁹ M,10⁻⁹ M, 5×10⁻¹⁰ M, 10⁻¹⁰ M, 5×10⁻¹¹ M, 10⁻¹¹ M, 5×10⁻¹² M, 10⁻¹² M,5×10⁻¹³ M, or 10⁻¹³ M. Preferably, the anti-PCSK9 antibodies andfragments thereof bind PCSK9 with a dissociation constant of less thanor equal to 10⁻¹¹ M, 5×10⁻¹² M, or 10⁻¹² M. In another embodiment of theinvention, the anti-PCSK9 antibodies of the present invention, andfragments thereof having binding specificity to PCSK9, bind to a linearor conformational PCSK9 epitope.

In another embodiment of the invention, the anti-PCSK9 activity of theanti-PCSK9 antibodies of the present invention, and fragments thereofhaving binding specificity to PCSK9, bind to PCSK9 with an off-rate ofless than or equal to 10⁻⁴ S⁻¹, 5×10⁻⁵ S⁻¹, 10⁻⁵ S⁻¹, 5×10⁻⁶ S⁻¹, 10⁻⁶S⁻¹, 5×10⁻⁷ S⁻¹, or 10⁻⁷ S⁻¹.

In a further embodiment of the invention, the anti-PCSK9 activity of theanti-PCSK9 antibodies of the present invention, and fragments thereofhaving binding specificity to PCSK9, exhibit anti-PCSK9 activity bypreventing, ameliorating or reducing the symptoms of, or alternativelytreating, diseases and disorders associated with PCSK9. Non-limitingexamples of diseases and disorders associated with PCSK9 are set forthherein.

Polynucleotides Encoding Anti-PCSK9 Antibody Polypeptides

Antibody Ab1

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 1:

(SEQ ID NO: 11) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtagctactggatgacttgggtccgccaggctccagggaaggggctggaatacatcggaatcattagtagtagtggtagcacatactacgcgacctgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtggatctggaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagagactctgcttttagttctggtttggaattcaacatctggggcccgggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagc ctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 2:

(SEQ ID NO: 12) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtagctactggatgacttgggtccgccaggctccagggaaggggctggaatacatcggaatcattagtagtagtggtagcacatactacgcgacctgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtggatctggaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagagactctgcttttagttctggtttggaattcaacatctggggcccgggcaccctcgtca ccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 10:

(SEQ ID NO: 20) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 21:

(SEQ ID NO: 31) gcctatgatctgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagtgtttatagtaactggttatcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgatgcatccgatctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtgcagtgtgacgatgctgccacttactactgtcagcaggggcagagtagtagtgatattgataatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagct tcaacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 22:

(SEQ ID NO: 32) gcctatgatctgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagtgtttatagtaactggttatcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgatgcatccgatctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcggcgtgcagtgtgacgatgctgccacttactactgtcagcaggggcagagtagtagtgatattgataatactttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 30:

(SEQ ID NO: 40) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 14; SEQ ID NO: 16; and SEQ ID NO: 18, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 1 orthe variable heavy chain sequence of SEQ ID NO: 2, and/or one or more ofthe polynucleotide sequences of SEQ ID NO: 34; SEQ ID NO: 36; and SEQ IDNO: 38, which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain sequence of SEQ IDNO: 21 or the variable light chain sequence of SEQ ID NO: 22, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 13; SEQ ID NO: 15; SEQ ID NO: 17; and SEQ ID NO: 19, whichcorrespond to polynucleotides encoding the framework regions (FRs orconstant regions) of the heavy chain sequence of SEQ ID NO: 1 or thevariable heavy chain sequence of SEQ ID NO: 2, and/or one or more of thepolynucleotide sequences of SEQ ID NO: 33; SEQ ID NO: 35; SEQ ID NO: 37;and SEQ ID NO: 39, which correspond to the framework regions (FRs orconstant regions) of the light chain sequence of SEQ ID NO: 21 or thevariable light chain sequence of SEQ ID NO: 22, or combinations of thesepolynucleotide sequences. In another embodiment of the invention, thepolynucleotides encoding the antibodies of the invention or fragmentsthereof comprise, or alternatively consist of, combinations of one ormore of the FRs, the variable heavy chain and variable light chainsequences, and the heavy chain and light chain sequences set forthabove, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 11 encoding the heavy chainsequence of SEQ ID NO: 1; the polynucleotide SEQ ID NO: 12 encoding thevariable heavy chain sequence of SEQ ID NO: 2; the polynucleotide SEQ IDNO: 31 encoding the light chain sequence of SEQ ID NO: 21; thepolynucleotide SEQ ID NO: 32 encoding the variable light chain sequenceof SEQ ID NO: 22; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 14; SEQ ID NO: 16; andSEQ ID NO: 18) of the heavy chain sequence of SEQ ID NO: 1 or thevariable heavy chain sequence of SEQ ID NO: 2; polynucleotides encodingthe complementarity-determining regions (SEQ ID NO: 34; SEQ ID NO: 36;and SEQ ID NO: 38) of the light chain sequence of SEQ ID NO: 21 or thevariable light chain sequence of SEQ ID NO: 22; polynucleotides encodingthe framework regions (SEQ ID NO: 13; SEQ ID NO: 15; SEQ ID NO: 17; andSEQ ID NO: 19) of the heavy chain sequence of SEQ ID NO: 1 or thevariable heavy chain sequence of SEQ ID NO: 2; and polynucleotidesencoding the framework regions (SEQ ID NO: 33; SEQ ID NO: 35; SEQ ID NO:37; and SEQ ID NO: 39) of the light chain sequence of SEQ ID NO: 21 orthe variable light chain sequence of SEQ ID NO: 22.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab1, the polynucleotidesencoding the full length Ab1 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 11 encoding the heavy chain sequenceof SEQ ID NO: 1 and the polynucleotide SEQ ID NO: 31 encoding the lightchain sequence of SEQ ID NO: 21.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab1 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab1 or Fab fragmentsthereof may be produced via expression of Ab1 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab2

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 41:

(SEQ ID NO: 51) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtagctatgcaatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcattgatgctattgataacacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatgaccagtctgacaaccggggacacggccacctatttctgtgccagagcctctattcttggttatagtattgctacgggctttaacatctggggcccagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 42:

(SEQ ID NO: 52) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtagctatgcaatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcattgatgctattgataacacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatgaccagtctgacaaccggggacacggccacctatttctgtgccagagcctctattcttggttatagtattgctacgggctttaacatctggggcccagggaccctcg tcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 50:

(SEQ ID NO: 60) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 61:

(SEQ ID NO: 71) gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtagccacttagcctggtatcagcagaaatcagggcagcctcccaagctcctgatctacagggcatccactctggaatctggggtctcatcaaggttcaaaggcagtggatctgggacagagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaacagggttatggtgttagtgatgttgataatggtttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtg t.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 62:

(SEQ ID NO: 72) gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtagccacttagcctggtatcagcagaaatcagggcagcctcccaagctcctgatctacagggcatccactctggaatctggggtctcatcaaggttcaaaggcagtggatctgggacagagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaacagggttatggtgttagtgatgttgataatggtttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 70:

(SEQ ID NO: 80) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 54; SEQ ID NO: 56; and SEQ ID NO: 58, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 41or the variable heavy chain sequence of SEQ ID NO: 42, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 74; SEQ ID NO: 76;and SEQ ID NO: 78, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 61 or the variable light chain sequence of SEQ ID NO: 62, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 53; SEQ ID NO: 55; SEQ ID NO: 57; and SEQ ID NO: 59, whichcorrespond to polynucleotides encoding the framework regions (FRs orconstant regions) of the heavy chain sequence of SEQ ID NO: 41 or thevariable heavy chain sequence of SEQ ID NO: 42, and/or one or more ofthe polynucleotide sequences of SEQ ID NO: 73; SEQ ID NO: 75; SEQ ID NO:77; and SEQ ID NO: 79, which correspond to the framework regions (FRs orconstant regions) of the light chain sequence of SEQ ID NO: 61 or thevariable light chain sequence of SEQ ID NO: 62, or combinations of thesepolynucleotide sequences. In another embodiment of the invention, thepolynucleotides encoding the antibodies of the invention or fragmentsthereof comprise, or alternatively consist of, combinations of one ormore of the FRs, the variable heavy chain and variable light chainsequences, and the heavy chain and light chain sequences set forthabove, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 51 encoding the heavy chainsequence of SEQ ID NO: 41; the polynucleotide SEQ ID NO: 52 encoding thevariable heavy chain sequence of SEQ ID NO: 42; the polynucleotide SEQID NO: 71 encoding the light chain sequence of SEQ ID NO: 61; thepolynucleotide SEQ ID NO: 72 encoding the variable light chain sequenceof SEQ ID NO: 62; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 54; SEQ ID NO: 56; andSEQ ID NO: 58) of the heavy chain sequence of SEQ ID NO: 41 or thevariable heavy chain sequence of SEQ ID NO: 42; polynucleotides encodingthe complementarity-determining regions (SEQ ID NO: 74; SEQ ID NO: 76;and SEQ ID NO: 78) of the light chain sequence of SEQ ID NO: 61 or thevariable light chain sequence of SEQ ID NO: 62; polynucleotides encodingthe framework regions (SEQ ID NO: 53; SEQ ID NO: 55; SEQ ID NO: 57; andSEQ ID NO: 59) of the heavy chain sequence of SEQ ID NO: 41 or thevariable heavy chain sequence of SEQ ID NO: 42; and polynucleotidesencoding the framework regions (SEQ ID NO: 73; SEQ ID NO: 75; SEQ ID NO:77; and SEQ ID NO: 79) of the light chain sequence of SEQ ID NO: 61 orthe variable light chain sequence of SEQ ID NO: 62.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab2, the polynucleotidesencoding the full length Ab2 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 51 encoding the heavy chain sequenceof SEQ ID NO: 41 and the polynucleotide SEQ ID NO: 71 encoding the lightchain sequence of SEQ ID NO: 61.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab2 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab2 or Fab fragmentsthereof may be produced via expression of Ab2 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab3

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 81:

(SEQ ID NO: 91) cagtcggtggaggagtccgggggtcgcctggtcacgcctggaggatccctgacactcacctgcacagcctctggattctccctcagtagctactacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttatcctagtggtagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacagtcgaggacacggccacctatttctgtgccagaggaggtgcttatgctactcttaacttgtggggcccgggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 82:

(SEQ ID NO: 92) cagtcggtggaggagtccgggggtcgcctggtcacgcctggaggatccctgacactcacctgcacagcctctggattctccctcagtagctactacatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcatttatcctagtggtagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacagtcgaggacacggccacctatttctgtgccagaggaggtgcttatgctactcttaacttgtggggcccgggcaccctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 90:

(SEQ ID NO: 100) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:101:

(SEQ ID NO: 111) gccgtgctgacccagacaccatcacccgtgtctgcagctgtgggaggcacagtcaccatcagttgccagtccagtcagagtgtttatcataacaacctcttatcctggtatcagcagaaaccaggtcagcctcccaagctcttgatctacgatgcatccaaactgacatctggggtctcatcgcggttcagcggcagtggatctgggacacagttcactctcaccataagcggcgtgcagtgtgacgatgctgccacttactactgtctaggcggttatgatgatgatgctgataatggtttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtg t.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 102:

(SEQ ID NO: 112) gccgtgctgacccagacaccatcacccgtgtctgcagctgtgggaggcacagtcaccatcagttgccagtccagtcagagtgtttatcataacaacctcttatcctggtatcagcagaaaccaggtcagcctcccaagctcttgatctacgatgcatccaaactgacatctggggtctcatcgcggttcagcggcagtggatctgggacacagttcactctcaccataagcggcgtgcagtgtgacgatgctgccacttactactgtctaggcggttatgatgatgatgctgataatggtttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 110:

(SEQ ID NO: 120) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 94; SEQ ID NO: 96; and SEQ ID NO: 98, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 81or the variable heavy chain sequence of SEQ ID NO: 82, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 114; SEQ ID NO: 116;and SEQ ID NO: 118, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 101 or the variable light chain sequence of SEQ ID NO: 102,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 93; SEQ ID NO: 95; SEQ ID NO: 97; and SEQ ID NO: 99, whichcorrespond to polynucleotides encoding the framework regions (FRs orconstant regions) of the heavy chain sequence of SEQ ID NO: 81 or thevariable heavy chain sequence of SEQ ID NO: 82, and/or one or more ofthe polynucleotide sequences of SEQ ID NO: 113; SEQ ID NO: 115; SEQ IDNO: 117; and SEQ ID NO: 119, which correspond to the framework regions(FRs or constant regions) of the light chain sequence of SEQ ID NO: 101or the variable light chain sequence of SEQ ID NO: 102, or combinationsof these polynucleotide sequences. In another embodiment of theinvention, the polynucleotides encoding the antibodies of the inventionor fragments thereof comprise, or alternatively consist of, combinationsof one or more of the FRs, the variable heavy chain and variable lightchain sequences, and the heavy chain and light chain sequences set forthabove, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 91 encoding the heavy chainsequence of SEQ ID NO: 81; the polynucleotide SEQ ID NO: 92 encoding thevariable heavy chain sequence of SEQ ID NO: 82; the polynucleotide SEQID NO: 111 encoding the light chain sequence of SEQ ID NO: 101; thepolynucleotide SEQ ID NO: 112 encoding the variable light chain sequenceof SEQ ID NO: 102; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 94; SEQ ID NO: 96; andSEQ ID NO: 98) of the heavy chain sequence of SEQ ID NO: 81 or thevariable heavy chain sequence of SEQ ID NO: 82; polynucleotides encodingthe complementarity-determining regions (SEQ ID NO: 114; SEQ ID NO: 116;and SEQ ID NO: 118) of the light chain sequence of SEQ ID NO: 101 or thevariable light chain sequence of SEQ ID NO: 102; polynucleotidesencoding the framework regions (SEQ ID NO: 93; SEQ ID NO: 95; SEQ ID NO:97; and SEQ ID NO: 99) of the heavy chain sequence of SEQ ID NO: 81 orthe variable heavy chain sequence of SEQ ID NO: 82; and polynucleotidesencoding the framework regions (SEQ ID NO: 113; SEQ ID NO: 115; SEQ IDNO: 117; and SEQ ID NO: 119) of the light chain sequence of SEQ ID NO:101 or the variable light chain sequence of SEQ ID NO: 102.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab3, the polynucleotidesencoding the full length Ab3 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 91 encoding the heavy chain sequenceof SEQ ID NO: 81 and the polynucleotide SEQ ID NO: 111 encoding thelight chain sequence of SEQ ID NO: 101.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab3 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab3 or Fab fragmentsthereof may be produced via expression of Ab3 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab4

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 121:

(SEQ ID NO: 131) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggaatcgacctcagtagctatgcaatgatctgggtccgtcaggctccagaaaaggggctggaatacatcggatatattggtggtattgatagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatgaccagtccgacaaccgaggacacggccacctatttctgtggcagatggtccggtactagtggttataataccatctggggcccgggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 122:

(SEQ ID NO: 132) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggaatcgacctcagtagctatgcaatgatctgggtccgtcaggctccagaaaaggggctggaatacatcggatatattggtggtattgatagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatgaccagtccgacaaccgaggacacggccacctatttctgtggcagatggtccggtactagtggttataataccatctggggcccgggcaccctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 130:

(SEQ ID NO: 140) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:141:

(SEQ ID NO: 151) gatgttgtgatgacccagactccagcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcatttatagcaatttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccaatctggcatctggggtctcatcgcggttcaaaggcagtcgatctgggacagagtacactctcaccatcagtgacctggagtgtgccgatgctgccacctactactgtcagtgcactggtggtggtgatagcggtaatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 142:

(SEQ ID NO: 152) gatgttgtgatgacccagactccagcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcatttatagcaatttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccaatctggcatctggggtctcatcgcggttcaaaggcagtcgatctgggacagagtacactctcaccatcagtgacctggagtgtgccgatgctgccacctactactgtcagtgcactggtggtggtgatagcggtaatactttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 150:

(SEQ ID NO: 160) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 134; SEQ ID NO: 136; and SEQ ID NO: 138, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 121or the variable heavy chain sequence of SEQ ID NO: 122, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 154; SEQ ID NO: 156;and SEQ ID NO: 158, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 141 or the variable light chain sequence of SEQ ID NO: 142,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 133; SEQ ID NO: 135; SEQ ID NO: 137; and SEQ ID NO: 139,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 121 orthe variable heavy chain sequence of SEQ ID NO: 122, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 153; SEQ ID NO: 155; SEQID NO: 157; and SEQ ID NO: 159, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 141 or the variable light chain sequence of SEQ ID NO: 142, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 131 encoding the heavy chainsequence of SEQ ID NO: 121; the polynucleotide SEQ ID NO: 132 encodingthe variable heavy chain sequence of SEQ ID NO: 122; the polynucleotideSEQ ID NO: 151 encoding the light chain sequence of SEQ ID NO: 141; thepolynucleotide SEQ ID NO: 152 encoding the variable light chain sequenceof SEQ ID NO: 142; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 134; SEQ ID NO: 136; andSEQ ID NO: 138) of the heavy chain sequence of SEQ ID NO: 121 or thevariable heavy chain sequence of SEQ ID NO: 122; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 154; SEQ IDNO: 156; and SEQ ID NO: 158) of the light chain sequence of SEQ ID NO:141 or the variable light chain sequence of SEQ ID NO: 142;polynucleotides encoding the framework regions (SEQ ID NO: 133; SEQ IDNO: 135; SEQ ID NO: 137; and SEQ ID NO: 139) of the heavy chain sequenceof SEQ ID NO: 121 or the variable heavy chain sequence of SEQ ID NO:122; and polynucleotides encoding the framework regions (SEQ ID NO: 153;SEQ ID NO: 155; SEQ ID NO: 157; and SEQ ID NO: 159) of the light chainsequence of SEQ ID NO: 141 or the variable light chain sequence of SEQID NO: 142.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab4, the polynucleotidesencoding the full length Ab4 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 131 encoding the heavy chain sequenceof SEQ ID NO: 121 and the polynucleotide SEQ ID NO: 151 encoding thelight chain sequence of SEQ ID NO: 141.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab4 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab4 or Fab fragmentsthereof may be produced via expression of Ab4 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab5

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 161:

(SEQ ID NO: 171) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtagctatgcaatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcattagtaatagtggtaccacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagaggaatatattggtactggagagtttttaacttgtggggcccggggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 162:

(SEQ ID NO: 172) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtagctatgcaatgagctgggtccgccaggctccagggaaggggctggaatggatcggaatcattagtaatagtggtaccacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacaaccgaggacacggccacctatttctgtgccagaggaatatattggtactggagagtttttaacttgtggggcccggggaccctcgtcaccgtc tcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 170:

(SEQ ID NO: 180) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:181:

(SEQ ID NO: 191) gccgtgctgacccagacaccatcgcctgtgtctgcagctgtgggaggcacagtcaccatcaattgccaggccagtcagagtgtttataacaacctcttatcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgatgcatccaatctggcatctggggtcccagataggttcagcggcagtggatctgggacacagttcactctcaccatcagcggcgtgcagtgtgacgatgctgccacttactactgtctaggcggttatgatgatgatgctgataatgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagctt caacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 182:

(SEQ ID NO: 192) gccgtgctgacccagacaccatcgcctgtgtctgcagctgtgggaggcacagtcaccatcaattgccaggccagtcagagtgtttataacaacctcttatcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgatgcatccaatctggcatctggggtcccagataggttcagcggcagtggatctgggacacagttcactctcaccatcagcggcgtgcagtgtgacgatgctgccacttactactgtctaggcggttatgatgatgatgctgataatgctttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 190:

(SEQ ID NO: 200) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 174; SEQ ID NO: 176; and SEQ ID NO: 178, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 161or the variable heavy chain sequence of SEQ ID NO: 162, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 194; SEQ ID NO: 196;and SEQ ID NO: 198, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 181 or the variable light chain sequence of SEQ ID NO: 182,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 173; SEQ ID NO: 175; SEQ ID NO: 177; and SEQ ID NO: 179,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 161 orthe variable heavy chain sequence of SEQ ID NO: 162, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 193; SEQ ID NO: 195; SEQID NO: 197; and SEQ ID NO: 199, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 181 or the variable light chain sequence of SEQ ID NO: 182, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 171 encoding the heavy chainsequence of SEQ ID NO: 161; the polynucleotide SEQ ID NO: 172 encodingthe variable heavy chain sequence of SEQ ID NO: 162; the polynucleotideSEQ ID NO: 191 encoding the light chain sequence of SEQ ID NO: 181; thepolynucleotide SEQ ID NO: 192 encoding the variable light chain sequenceof SEQ ID NO: 182; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 174; SEQ ID NO: 176; andSEQ ID NO: 178) of the heavy chain sequence of SEQ ID NO: 161 or thevariable heavy chain sequence of SEQ ID NO: 162; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 194; SEQ IDNO: 196; and SEQ ID NO: 198) of the light chain sequence of SEQ ID NO:181 or the variable light chain sequence of SEQ ID NO: 182;polynucleotides encoding the framework regions (SEQ ID NO: 173; SEQ IDNO: 175; SEQ ID NO: 177; and SEQ ID NO: 179) of the heavy chain sequenceof SEQ ID NO: 161 or the variable heavy chain sequence of SEQ ID NO:162; and polynucleotides encoding the framework regions (SEQ ID NO: 193;SEQ ID NO: 195; SEQ ID NO: 197; and SEQ ID NO: 199) of the light chainsequence of SEQ ID NO: 181 or the variable light chain sequence of SEQID NO: 182.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab5, the polynucleotidesencoding the full length Ab5 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 171 encoding the heavy chain sequenceof SEQ ID NO: 161 and the polynucleotide SEQ ID NO: 191 encoding thelight chain sequence of SEQ ID NO: 181.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab5 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab5 or Fab fragmentsthereof may be produced via expression of Ab5 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab6

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 201:

(SEQ ID NO: 211) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacagcctctggattctccttcagtagcaactactggatatgctgggtccgccaggctccagggaagggactggagtggatcggatgcattcgtgatggtggtggcacttactacgcgagctgggcgaaaggccgactcaccatctccatgacctcgtcgaccacggtgactctgcaactgaacagtctgacagccgcggacacggccacctatttttgtgcgagcgatattaatgatgggtggcttggccaattcaacttgtggggcccaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 202:

(SEQ ID NO: 212) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacagcctctggattctccttcagtagcaactactggatatgctgggtccgccaggctccagggaagggactggagtggatcggatgcattcgtgatggtggtggcacttactacgcgagctgggcgaaaggccgactcaccatctccatgacctcgtcgaccacggtgactctgcaactgaacagtctgacagccgcggacacggccacctatttttgtgcgagcgatattaatgatgggtggcttggccaattcaacttgtggggcccaggcacc ctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 210:

(SEQ ID NO: 220) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccct gtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:221:

(SEQ ID NO: 231) gctgacattgtgatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtgcgtacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacagggcatacactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactaatacttatggaaatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 222:

(SEQ ID NO: 232) gctgacattgtgatgacccagactccagcctctgtggaggtagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtgcgtacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacagggcatacactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactaatacttatggaaatactttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 230:

(SEQ ID NO: 240) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 214; SEQ ID NO: 216; and SEQ ID NO: 218, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 201or the variable heavy chain sequence of SEQ ID NO: 202, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 234; SEQ ID NO: 236;and SEQ ID NO: 238, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 221 or the variable light chain sequence of SEQ ID NO: 222,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 213; SEQ ID NO: 215; SEQ ID NO: 217; and SEQ ID NO: 219,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 201 orthe variable heavy chain sequence of SEQ ID NO: 202, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 233; SEQ ID NO: 235; SEQID NO: 237; and SEQ ID NO: 239, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 221 or the variable light chain sequence of SEQ ID NO: 222, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 211 encoding the heavy chainsequence of SEQ ID NO: 201; the polynucleotide SEQ ID NO: 212 encodingthe variable heavy chain sequence of SEQ ID NO: 202; the polynucleotideSEQ ID NO: 231 encoding the light chain sequence of SEQ ID NO: 221; thepolynucleotide SEQ ID NO: 232 encoding the variable light chain sequenceof SEQ ID NO: 222; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 214; SEQ ID NO: 216; andSEQ ID NO: 218) of the heavy chain sequence of SEQ ID NO: 201 or thevariable heavy chain sequence of SEQ ID NO: 202; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 234; SEQ IDNO: 236; and SEQ ID NO: 238) of the light chain sequence of SEQ ID NO:221 or the variable light chain sequence of SEQ ID NO: 222;polynucleotides encoding the framework regions (SEQ ID NO: 213; SEQ IDNO: 215; SEQ ID NO: 217; and SEQ ID NO: 219) of the heavy chain sequenceof SEQ ID NO: 201 or the variable heavy chain sequence of SEQ ID NO:202; and polynucleotides encoding the framework regions (SEQ ID NO: 233;SEQ ID NO: 235; SEQ ID NO: 237; and SEQ ID NO: 239) of the light chainsequence of SEQ ID NO: 221 or the variable light chain sequence of SEQID NO: 222.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab6, the polynucleotidesencoding the full length Ab6 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 211 encoding the heavy chain sequenceof SEQ ID NO: 201 and the polynucleotide SEQ ID NO: 231 encoding thelight chain sequence of SEQ ID NO: 221.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab6 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab6 or Fab fragmentsthereof may be produced via expression of Ab6 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab7

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 241:

(SEQ ID NO: 251) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaactactggatatgctgggtccgtcaggctccagggaaggggctggagtggatcggatgcattcgtgatggtggtggcacttactacgctagctctgctaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagcgatatcaatgatgggtggcttggccaattcaacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggagcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 242:

(SEQ ID NO: 252) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaactactggatatgctgggtccgtcaggctccagggaaggggctggagtggatcggatgcattcgtgatggtggtggcacttactacgctagctctgctaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagcgatatcaatgatgggtggcttggccaattcaacttgtggggccaaggga ccctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 250:

(SEQ ID NO: 260) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:261:

(SEQ ID NO: 271) gctgacattgtgatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcaagtgccaggccagtcagagcattagtgcttacttagcctggtatcagcagaaaccagggaaagtccctaagctcctgatctatagggcatacactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaaagctactattccgttactactaatacttatggaaatactttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacag gggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 262:

(SEQ ID NO: 272) gctgacattgtgatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcaagtgccaggccagtcagagcattagtgcttacttagcctggtatcagcagaaaccagggaaagtccctaagctcctgatctatagggcatacactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaaagctactattccgttactactaatacttatggaaatactttcggcggaggaaccaaggtggaaatcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 270:

(SEQ ID NO: 280) cgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 254; SEQ ID NO: 256; and SEQ ID NO: 258, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 241or the variable heavy chain sequence of SEQ ID NO: 242, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 274; SEQ ID NO: 276;and SEQ ID NO: 278, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 261 or the variable light chain sequence of SEQ ID NO: 262,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 253; SEQ ID NO: 255; SEQ ID NO: 257; and SEQ ID NO: 259,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 241 orthe variable heavy chain sequence of SEQ ID NO: 242, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 273; SEQ ID NO: 275; SEQID NO: 277; and SEQ ID NO: 279, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 261 or the variable light chain sequence of SEQ ID NO: 262, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 251 encoding the heavy chainsequence of SEQ ID NO: 241; the polynucleotide SEQ ID NO: 252 encodingthe variable heavy chain sequence of SEQ ID NO: 242; the polynucleotideSEQ ID NO: 271 encoding the light chain sequence of SEQ ID NO: 261; thepolynucleotide SEQ ID NO: 272 encoding the variable light chain sequenceof SEQ ID NO: 262; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 254; SEQ ID NO: 256; andSEQ ID NO: 258) of the heavy chain sequence of SEQ ID NO: 241 or thevariable heavy chain sequence of SEQ ID NO: 242; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 274; SEQ IDNO: 276; and SEQ ID NO: 278) of the light chain sequence of SEQ ID NO:261 or the variable light chain sequence of SEQ ID NO: 262;polynucleotides encoding the framework regions (SEQ ID NO: 253; SEQ IDNO: 255; SEQ ID NO: 257; and SEQ ID NO: 259) of the heavy chain sequenceof SEQ ID NO: 241 or the variable heavy chain sequence of SEQ ID NO:242; and polynucleotides encoding the framework regions (SEQ ID NO: 273;SEQ ID NO: 275; SEQ ID NO: 277; and SEQ ID NO: 279) of the light chainsequence of SEQ ID NO: 261 or the variable light chain sequence of SEQID NO: 262.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab7, the polynucleotidesencoding the full length Ab7 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 251 encoding the heavy chain sequenceof SEQ ID NO: 241 and the polynucleotide SEQ ID NO: 271 encoding thelight chain sequence of SEQ ID NO: 261.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab7 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab7 or Fab fragmentsthereof may be produced via expression of Ab7 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab8

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 281:

(SEQ ID NO: 291) caggagcagctggtggagtccgggggaggcctggtccagcctgagggatccctgacactcacctgcacagcttctggattctccttcactagcgactattacatgtgctgggtccgccaggctccagggaaggggctggagtggatcggatgcatttctactggtgatggcagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaccctcgtcgaccacggtgactctgcaaatgaccaggctgacagccgcggacacggccacctatttctgtgcgagagatcgatactatagttatgcttatggtgcttatgtttatgctagcgacttgtggggcccaggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctc tccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 282:

(SEQ ID NO: 292) caggagcagctggtggagtccgggggaggcctggtccagcctgagggatccctgacactcacctgcacagcttctggattctccttcactagcgactattacatgtgctgggtccgccaggctccagggaaggggctggagtggatcggatgcatttctactggtgatggcagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaccctcgtcgaccacggtgactctgcaaatgaccaggctgacagccgcggacacggccacctatttctgtgcgagagatcgatactatagttatgcttatggtgcttatgtttatgctagcgacttgtggggcccaggcaccctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 290:

(SEQ ID NO: 300) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:301:

(SEQ ID NO: 311) gctgacattgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaattgccaggccagtgaaagcattaggaactacttatcctggtatcaacagaaaccagggcagcgtcccaagctcctgatctatggtgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagatttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagcaattatggtattagtagtcgtagttatgttaatggtttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacag gggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 302:

(SEQ ID NO: 312) gctgacattgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaattgccaggccagtgaaagcattaggaactacttatcctggtatcaacagaaaccagggcagcgtcccaagctcctgatctatggtgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagatttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagcaattatggtattagtagtcgtagttatgttaatggtttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 310:

(SEQ ID NO: 320) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 294; SEQ ID NO: 296; and SEQ ID NO: 298, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 281or the variable heavy chain sequence of SEQ ID NO: 282, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 314; SEQ ID NO: 316;and SEQ ID NO: 318, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 301 or the variable light chain sequence of SEQ ID NO: 302,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 293; SEQ ID NO: 295; SEQ ID NO: 297; and SEQ ID NO: 299,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 281 orthe variable heavy chain sequence of SEQ ID NO: 282, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 313; SEQ ID NO: 315; SEQID NO: 317; and SEQ ID NO: 319, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 301 or the variable light chain sequence of SEQ ID NO: 302, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 291 encoding the heavy chainsequence of SEQ ID NO: 281; the polynucleotide SEQ ID NO: 292 encodingthe variable heavy chain sequence of SEQ ID NO: 282; the polynucleotideSEQ ID NO: 311 encoding the light chain sequence of SEQ ID NO: 301; thepolynucleotide SEQ ID NO: 312 encoding the variable light chain sequenceof SEQ ID NO: 302; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 294; SEQ ID NO: 296; andSEQ ID NO: 298) of the heavy chain sequence of SEQ ID NO: 281 or thevariable heavy chain sequence of SEQ ID NO: 282; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 314; SEQ IDNO: 316; and SEQ ID NO: 318) of the light chain sequence of SEQ ID NO:301 or the variable light chain sequence of SEQ ID NO: 302;polynucleotides encoding the framework regions (SEQ ID NO: 293; SEQ IDNO: 295; SEQ ID NO: 297; and SEQ ID NO: 299) of the heavy chain sequenceof SEQ ID NO: 281 or the variable heavy chain sequence of SEQ ID NO:282; and polynucleotides encoding the framework regions (SEQ ID NO: 313;SEQ ID NO: 315; SEQ ID NO: 317; and SEQ ID NO: 319) of the light chainsequence of SEQ ID NO: 301 or the variable light chain sequence of SEQID NO: 302.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab8, the polynucleotidesencoding the full length Ab8 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 291 encoding the heavy chain sequenceof SEQ ID NO: 281 and the polynucleotide SEQ ID NO: 311 encoding thelight chain sequence of SEQ ID NO: 301.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab8 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab8 or Fab fragmentsthereof may be produced via expression of Ab8 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab9

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 321:

(SEQ ID NO: 331) cagtcggtggaggagtccgggggtcgcctggtaacgcctgggacacccctgacactcacctgcacagtctctggaatcgacctcagtagctatgcaatgggctgggtccgccaggctccagggaaggggctggaatacatcggaatcattgttagttatgggcccacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacggccgaggacacggccacctatttctgtgccagagatctggatgctaatagtagtggttattatggatgctttaacatctggggccaggggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 322:

(SEQ ID NO: 332) cagtcggtggaggagtccgggggtcgcctggtaacgcctgggacacccctgacactcacctgcacagtctctggaatcgacctcagtagctatgcaatgggctgggtccgccaggctccagggaaggggctggaatacatcggaatcattgttagttatgggcccacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacggccgaggacacggccacctatttctgtgccagagatctggatgctaatagtagtggttattatggatgctttaacatctggggccaggggaccctcgtcac cgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 330:

(SEQ ID NO: 340) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:341:

(SEQ ID NO: 351) gccgtcgtgctgacccagactccagcctccgtgtctgcagctgtgggtggcacagtcaccatcaagtgccaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgctgcatcccctctggcatctggggtctcatcgcggttcaagagcagtggatctgggacagagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttattactgtcaaagctattatggtagtagcaatattgctttcggcggagggaccgagctggagatcctacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 342:

(SEQ ID NO: 352) gccgtcgtgctgacccagactccagcctccgtgtctgcagctgtgggtggcacagtcaccatcaagtgccaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatgctgcatcccctctggcatctggggtctcatcgcggttcaagagcagtggatctgggacagagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttattactgtcaaagctattatggtagtagcaatattgctttcggcggagggaccgagctggagatccta.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 350:

(SEQ ID NO: 360) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 334; SEQ ID NO: 336; and SEQ ID NO: 338, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 321or the variable heavy chain sequence of SEQ ID NO: 322, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 354; SEQ ID NO: 356;and SEQ ID NO: 358, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 341 or the variable light chain sequence of SEQ ID NO: 342,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 333; SEQ ID NO: 335; SEQ ID NO: 337; and SEQ ID NO: 339,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 321 orthe variable heavy chain sequence of SEQ ID NO: 322, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 353; SEQ ID NO: 355; SEQID NO: 357; and SEQ ID NO: 359, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 341 or the variable light chain sequence of SEQ ID NO: 342, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 331 encoding the heavy chainsequence of SEQ ID NO: 321; the polynucleotide SEQ ID NO: 332 encodingthe variable heavy chain sequence of SEQ ID NO: 322; the polynucleotideSEQ ID NO: 351 encoding the light chain sequence of SEQ ID NO: 341; thepolynucleotide SEQ ID NO: 352 encoding the variable light chain sequenceof SEQ ID NO: 342; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 334; SEQ ID NO: 336; andSEQ ID NO: 338) of the heavy chain sequence of SEQ ID NO: 321 or thevariable heavy chain sequence of SEQ ID NO: 322; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 354; SEQ IDNO: 356; and SEQ ID NO: 358) of the light chain sequence of SEQ ID NO:341 or the variable light chain sequence of SEQ ID NO: 342;polynucleotides encoding the framework regions (SEQ ID NO: 333; SEQ IDNO: 335; SEQ ID NO: 337; and SEQ ID NO: 339) of the heavy chain sequenceof SEQ ID NO: 321 or the variable heavy chain sequence of SEQ ID NO:322; and polynucleotides encoding the framework regions (SEQ ID NO: 353;SEQ ID NO: 355; SEQ ID NO: 357; and SEQ ID NO: 359) of the light chainsequence of SEQ ID NO: 341 or the variable light chain sequence of SEQID NO: 342.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab9, the polynucleotidesencoding the full length Ab9 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 331 encoding the heavy chain sequenceof SEQ ID NO: 321 and the polynucleotide SEQ ID NO: 351 encoding thelight chain sequence of SEQ ID NO: 341.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab9 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab9 or Fab fragmentsthereof may be produced via expression of Ab9 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab10

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 361:

(SEQ ID NO: 371) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacagcctctggattctccttcagtagcagttactggatatgctgggtccgccaggctccagggaaggggctggagtggatcgcatgcattcgtgctggtggtgggaattactacgcgaactgggcgaaaggccgattcaccatctccagaacctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttttgtgcgagcgatattaatgatgggtggcttggccaattcaacttgtggggcccgggcaccctggtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 362:

(SEQ ID NO: 372) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacagcctctggattctccttcagtagcagttactggatatgctgggtccgccaggctccagggaaggggctggagtggatcgcatgcattcgtgctggtggtgggaattactacgcgaactgggcgaaaggccgattcaccatctccagaacctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttttgtgcgagcgatattaatgatgggtggcttggccaattcaacttgtggggcccgggcaccctggtcac cgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 370:

(SEQ ID NO: 380) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:381:

(SEQ ID NO: 391) gccaacattgtgatgacccagactccagcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtaattacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacaggacatccactctggcatctggggtcccatcgcggttcaaaggcagtggatccgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactgttgcttatggaaatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacag gggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 382:

(SEQ ID NO: 392) gccaacattgtgatgacccagactccagcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtaattacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacaggacatccactctggcatctggggtcccatcgcggttcaaaggcagtggatccgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactgttgcttatggaaatactttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 390:

(SEQ ID NO: 400) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 374; SEQ ID NO: 376; and SEQ ID NO: 378, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 361or the variable heavy chain sequence of SEQ ID NO: 362, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 394; SEQ ID NO: 396;and SEQ ID NO: 398, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 381 or the variable light chain sequence of SEQ ID NO: 382,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 373; SEQ ID NO: 375; SEQ ID NO: 377; and SEQ ID NO: 379,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 361 orthe variable heavy chain sequence of SEQ ID NO: 362, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 393; SEQ ID NO: 395; SEQID NO: 397; and SEQ ID NO: 399, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 381 or the variable light chain sequence of SEQ ID NO: 382, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 371 encoding the heavy chainsequence of SEQ ID NO: 361; the polynucleotide SEQ ID NO: 372 encodingthe variable heavy chain sequence of SEQ ID NO: 362; the polynucleotideSEQ ID NO: 391 encoding the light chain sequence of SEQ ID NO: 381; thepolynucleotide SEQ ID NO: 392 encoding the variable light chain sequenceof SEQ ID NO: 382; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 374; SEQ ID NO: 376; andSEQ ID NO: 378) of the heavy chain sequence of SEQ ID NO: 361 or thevariable heavy chain sequence of SEQ ID NO: 362; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 394; SEQ IDNO: 396; and SEQ ID NO: 398) of the light chain sequence of SEQ ID NO:381 or the variable light chain sequence of SEQ ID NO: 382;polynucleotides encoding the framework regions (SEQ ID NO: 373; SEQ IDNO: 375; SEQ ID NO: 377; and SEQ ID NO: 379) of the heavy chain sequenceof SEQ ID NO: 361 or the variable heavy chain sequence of SEQ ID NO:362; and polynucleotides encoding the framework regions (SEQ ID NO: 393;SEQ ID NO: 395; SEQ ID NO: 397; and SEQ ID NO: 399) of the light chainsequence of SEQ ID NO: 381 or the variable light chain sequence of SEQID NO: 382.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab10, thepolynucleotides encoding the full length Ab10 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 371 encoding theheavy chain sequence of SEQ ID NO: 361 and the polynucleotide SEQ ID NO:391 encoding the light chain sequence of SEQ ID NO: 381.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab10 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab10 or Fab fragmentsthereof may be produced via expression of Ab10 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab11

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 401:

(SEQ ID NO: 411) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcagttactggatatgctgggtccgtcaggctccagggaaggggctggagtggatcgcatgcattcgtgctggtggtgggaattactacgctaactctgctaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagcgatatcaatgatgggtggcttggccaattcaacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggt aaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 402:

(SEQ ID NO: 412) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcagttactggatatgctgggtccgtcaggctccagggaaggggctggagtggatcgcatgcattcgtgctggtggtgggaattactacgctaactctgctaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagcgatatcaatgatgggtggcttggccaattcaacttgtggggccaagggaccctcgt caccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 410:

(SEQ ID NO: 420) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaacttgacctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggcatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:421:

(SEQ ID NO: 431) gccaacattgtgatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattagtaattacttagcctggtatcagcagaaaccagggaaagtccctaagctcctgatctataggacatccactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaaagctactattccgttactactgttgcttatggaaatactttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacag gggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 422:

(SEQ ID NO: 432) gccaacattgtgatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgccaggccagtcagagcattagtaattacttagcctggtatcagcagaaaccagggaaagtccctaagctcctgatctataggacatccactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaaagctactattccgttactactgttgcttatggaaatactttcggcggaggaaccaaggtggaaatcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 430:

(SEQ ID NO: 440) cgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 414; SEQ ID NO: 416; and SEQ ID NO: 418, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 401or the variable heavy chain sequence of SEQ ID NO: 402, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 434; SEQ ID NO: 436;and SEQ ID NO: 438, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 421 or the variable light chain sequence of SEQ ID NO: 422,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 413; SEQ ID NO: 415; SEQ ID NO: 417; and SEQ ID NO: 419,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 401 orthe variable heavy chain sequence of SEQ ID NO: 402, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 433; SEQ ID NO: 435; SEQID NO: 437; and SEQ ID NO: 439, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 421 or the variable light chain sequence of SEQ ID NO: 422, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 411 encoding the heavy chainsequence of SEQ ID NO: 401; the polynucleotide SEQ ID NO: 412 encodingthe variable heavy chain sequence of SEQ ID NO: 402; the polynucleotideSEQ ID NO: 431 encoding the light chain sequence of SEQ ID NO: 421; thepolynucleotide SEQ ID NO: 432 encoding the variable light chain sequenceof SEQ ID NO: 422; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 414; SEQ ID NO: 416; andSEQ ID NO: 418) of the heavy chain sequence of SEQ ID NO: 401 or thevariable heavy chain sequence of SEQ ID NO: 402; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 434; SEQ IDNO: 436; and SEQ ID NO: 438) of the light chain sequence of SEQ ID NO:421 or the variable light chain sequence of SEQ ID NO: 422;polynucleotides encoding the framework regions (SEQ ID NO: 413; SEQ IDNO: 415; SEQ ID NO: 417; and SEQ ID NO: 419) of the heavy chain sequenceof SEQ ID NO: 401 or the variable heavy chain sequence of SEQ ID NO:402; and polynucleotides encoding the framework regions (SEQ ID NO: 433;SEQ ID NO: 435; SEQ ID NO: 437; and SEQ ID NO: 439) of the light chainsequence of SEQ ID NO: 421 or the variable light chain sequence of SEQID NO: 422.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab11, thepolynucleotides encoding the full length Ab11 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 411 encoding theheavy chain sequence of SEQ ID NO: 401 and the polynucleotide SEQ ID NO:431 encoding the light chain sequence of SEQ ID NO: 421.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab11 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab11 or Fab fragmentsthereof may be produced via expression of Ab11 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab12

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 441:

(SEQ ID NO: 451) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcagttactggatatgctgggtccgtcaggctccagggaaggggctggagtggatcgcatgcattcgtgctggtggtgggaattactacgctaactctgctaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagcgatatcaatgatgggtggcttggccaattcaacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggt aaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 442:

(SEQ ID NO: 452) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcagttactggatatgctgggtccgtcaggctccagggaaggggctggagtggatcgcatgcattcgtgctggtggtgggaattactacgctaactctgctaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagcgatatcaatgatgggtggcttggccaattcaacttgtggggccaagggaccctcgt caccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 450:

(SEQ ID NO: 460) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:461:

(SEQ ID NO: 471) gccaacattgtgatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcaagtgccaggccagtcagagcattagtaattacttagcctggtatcagcagaaaccagggaaagtccctaagctcctgatctataggacatccactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaaagctactattccgttactactgttgcttatggaaatactttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 462:

(SEQ ID NO: 472) gccaacattgtgatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcaagtgccaggccagtcagagcattagtaattacttagcctggtatcagcagaaaccagggaaagtccctaagctcctgatc  tataggacatccactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctga  agatgttgcaacttattactgtcaaagctactattccgttactactgttgcttatggaaatactttcggcggaggaaccaaggtggaaatcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 470:

(SEQ ID NO: 480) cgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 454; SEQ ID NO: 456; and SEQ ID NO: 458, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 441or the variable heavy chain sequence of SEQ ID NO: 442, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 474; SEQ ID NO: 476;and SEQ ID NO: 478, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 461 or the variable light chain sequence of SEQ ID NO: 462,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 453; SEQ ID NO: 455; SEQ ID NO: 457; and SEQ ID NO: 459,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 441 orthe variable heavy chain sequence of SEQ ID NO: 442, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 473; SEQ ID NO: 475; SEQID NO: 477; and SEQ ID NO: 479, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 461 or the variable light chain sequence of SEQ ID NO: 462, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 451 encoding the heavy chainsequence of SEQ ID NO: 441; the polynucleotide SEQ ID NO: 452 encodingthe variable heavy chain sequence of SEQ ID NO: 442; the polynucleotideSEQ ID NO: 471 encoding the light chain sequence of SEQ ID NO: 461; thepolynucleotide SEQ ID NO: 472 encoding the variable light chain sequenceof SEQ ID NO: 462; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 454; SEQ ID NO: 456; andSEQ ID NO: 458) of the heavy chain sequence of SEQ ID NO: 441 or thevariable heavy chain sequence of SEQ ID NO: 442; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 474; SEQ IDNO: 476; and SEQ ID NO: 478) of the light chain sequence of SEQ ID NO:461 or the variable light chain sequence of SEQ ID NO: 462;polynucleotides encoding the framework regions (SEQ ID NO: 453; SEQ IDNO: 455; SEQ ID NO: 457; and SEQ ID NO: 459) of the heavy chain sequenceof SEQ ID NO: 441 or the variable heavy chain sequence of SEQ ID NO:442; and polynucleotides encoding the framework regions (SEQ ID NO: 473;SEQ ID NO: 475; SEQ ID NO: 477; and SEQ ID NO: 479) of the light chainsequence of SEQ ID NO: 461 or the variable light chain sequence of SEQID NO: 462.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab12, thepolynucleotides encoding the full length Ab12 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 451 encoding theheavy chain sequence of SEQ ID NO: 441 and the polynucleotide SEQ ID NO:471 encoding the light chain sequence of SEQ ID NO: 461.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab12 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab12 or Fab fragmentsthereof may be produced via expression of Ab12 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab13

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 481:

(SEQ ID NO: 491) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggaatcgacctcagtacctatggagtgggctgggtccgccaggctccagggaaggggctggaatacatcggaatcattagtagtagtggtagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtggatctgaaaatgaccagtctgacaaccgaggacacggccacctatttctgtgccagagattggtctagtactactggttattatgggtattttaatatgtggggcccgggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 482:

(SEQ ID NO: 492) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccc  tgacactcacctgcacagtctctggaatcgacctcagtacctatggagtgggctgggtccgccaggctccagggaaggggctggaatacatcggaatc  attagtagtagtggtagcacatactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtggatctgaaaatgaccag  tctgacaaccgaggacacggccacctatttctgtgccagagattggtctagtactactggttattatgggtattttaatatgtggggcccgggcaccc tcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 490:

(SEQ ID NO: 500) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:501:

(SEQ ID NO: 511) gcattcgaattgacccagactccatcccccgtgtctgcagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccaatctggaatctggggtcccatcgcggttcagcggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccatttactactgtcaaagctcttatggtagtagtactttggctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggg gagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 502:

(SEQ ID NO: 512) gcattcgaattgacccagactccatcccccgtgtctgcagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccaatctggaatctggggtcccatcgcggttcagcggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccatttactactgtcaaagctcttatggtagtagtactttggctttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 510:

(SEQ ID NO: 520) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 494; SEQ ID NO: 496; and SEQ ID NO: 498, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 481or the variable heavy chain sequence of SEQ ID NO: 482, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 514; SEQ ID NO: 516;and SEQ ID NO: 518, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 501 or the variable light chain sequence of SEQ ID NO: 502,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 493; SEQ ID NO: 495; SEQ ID NO: 497; and SEQ ID NO: 499,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 481 orthe variable heavy chain sequence of SEQ ID NO: 482, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 513; SEQ ID NO: 515; SEQID NO: 517; and SEQ ID NO: 519, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 501 or the variable light chain sequence of SEQ ID NO: 502, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 491 encoding the heavy chainsequence of SEQ ID NO: 481; the polynucleotide SEQ ID NO: 492 encodingthe variable heavy chain sequence of SEQ ID NO: 482; the polynucleotideSEQ ID NO: 511 encoding the light chain sequence of SEQ ID NO: 501; thepolynucleotide SEQ ID NO: 512 encoding the variable light chain sequenceof SEQ ID NO: 502; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 494; SEQ ID NO: 496; andSEQ ID NO: 498) of the heavy chain sequence of SEQ ID NO: 481 or thevariable heavy chain sequence of SEQ ID NO: 482; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 514; SEQ IDNO: 516; and SEQ ID NO: 518) of the light chain sequence of SEQ ID NO:501 or the variable light chain sequence of SEQ ID NO: 502;polynucleotides encoding the framework regions (SEQ ID NO: 493; SEQ IDNO: 495; SEQ ID NO: 497; and SEQ ID NO: 499) of the heavy chain sequenceof SEQ ID NO: 481 or the variable heavy chain sequence of SEQ ID NO:482; and polynucleotides encoding the framework regions (SEQ ID NO: 513;SEQ ID NO: 515; SEQ ID NO: 517; and SEQ ID NO: 519) of the light chainsequence of SEQ ID NO: 501 or the variable light chain sequence of SEQID NO: 502.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab13, thepolynucleotides encoding the full length Ab13 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 491 encoding theheavy chain sequence of SEQ ID NO: 481 and the polynucleotide SEQ ID NO:511 encoding the light chain sequence of SEQ ID NO: 501.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab13 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab13 or Fab fragmentsthereof may be produced via expression of Ab13 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab14

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 521:

(SEQ ID NO: 531) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacaggttctggattctccttcagtagcatcgcctacatgtgctggatccgccaggctccagggaaggggctggagtggatcggatgcattggttctggtagtgggaacacttactacgcgaactgggcgaaaggccgattcaccatctccaaaagctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacttatttctgtgcgagcgatactaataatgggtggcttggccaattcaacttgtggggccagggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 522:

(SEQ ID NO: 532) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacaggttctggattctccttcagtagcatcgcctacatgtgctggatccgccaggctccagggaaggggctggagtggatcggatgcattggttctggtagtgggaacacttactacgcgaactgggcgaaaggccgattcaccatctccaaaagctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacttatttctgtgcgagcgatactaataatgggtggcttggccaattcaacttgtggggccagggca ccctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 530:

(SEQ ID NO: 540) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:541:

(SEQ ID NO: 551) gctgacattgtgatgacccagactccagcctcggtgtctgcagctgtgggaggcacagtcaccatcaattgccaggccagtcagagcattagtagctacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacagggcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaggctattattccgttactactaatacttatggaaatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 542:gctgacattgtgatgacccagactccagcctcggtgtctgcagctgtgggaggcacagtcaccatcaattgccaggccagtcagagcattagtagctacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacagggcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaggctattattccgttactactaatacttatggaaatactttcggcggagggaccgaggtggtggtcaaa(SEQ ID NO: 552).

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 550:

(SEQ ID NO: 560) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 534; SEQ ID NO: 536; and SEQ ID NO: 538, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 521or the variable heavy chain sequence of SEQ ID NO: 522, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 554; SEQ ID NO: 556;and SEQ ID NO: 558, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 541 or the variable light chain sequence of SEQ ID NO: 542,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 533; SEQ ID NO: 535; SEQ ID NO: 537; and SEQ ID NO: 539,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 521 orthe variable heavy chain sequence of SEQ ID NO: 522, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 553; SEQ ID NO: 555; SEQID NO: 557; and SEQ ID NO: 559, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 541 or the variable light chain sequence of SEQ ID NO: 542, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 531 encoding the heavy chainsequence of SEQ ID NO: 521; the polynucleotide SEQ ID NO: 532 encodingthe variable heavy chain sequence of SEQ ID NO: 522; the polynucleotideSEQ ID NO: 551 encoding the light chain sequence of SEQ ID NO: 541; thepolynucleotide SEQ ID NO: 552 encoding the variable light chain sequenceof SEQ ID NO: 542; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 534; SEQ ID NO: 536; andSEQ ID NO: 538) of the heavy chain sequence of SEQ ID NO: 521 or thevariable heavy chain sequence of SEQ ID NO: 522; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 554; SEQ IDNO: 556; and SEQ ID NO: 558) of the light chain sequence of SEQ ID NO:541 or the variable light chain sequence of SEQ ID NO: 542;polynucleotides encoding the framework regions (SEQ ID NO: 533; SEQ IDNO: 535; SEQ ID NO: 537; and SEQ ID NO: 539) of the heavy chain sequenceof SEQ ID NO: 521 or the variable heavy chain sequence of SEQ ID NO:522; and polynucleotides encoding the framework regions (SEQ ID NO: 553;SEQ ID NO: 555; SEQ ID NO: 557; and SEQ ID NO: 559) of the light chainsequence of SEQ ID NO: 541 or the variable light chain sequence of SEQID NO: 542.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab14, thepolynucleotides encoding the full length Ab14 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 531 encoding theheavy chain sequence of SEQ ID NO: 521 and the polynucleotide SEQ ID NO:551 encoding the light chain sequence of SEQ ID NO: 541.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab14 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab14 or Fab fragmentsthereof may be produced via expression of Ab14 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab15

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 561:

(SEQ ID NO: 571) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacagcctctggattctccttcagtagcagctactggatatgctgggtccgccaggctccagggaagggactggagtggatcgcatgcattgatgctggtaatagtggtagcacttactacgcgagctgggcgaaaggccgattcaccatctccaaggcctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttttgtgcgagcgatcttaatgatgggtggcttggccaattcaacttgtggggcccgggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 562:

(SEQ ID NO: 572) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacagcctctggattctccttcagtagcagctactggatatgctgggtccgccaggctccagggaagggactggagtggatcgcatgcattgatgctggtaatagtggtagcacttactacgcgagctgggcgaaaggccgattcaccatctccaaggcctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttttgtgcgagcgatcttaatgatgggtggcttggccaattcaacttgtggggcccgggcaccctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 570:

(SEQ ID NO: 580) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:581:

(SEQ ID NO: 591) gccaacatcgtgatgacccagactccatcccccgtgtctggagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtgactacttagcctggtatcagcagaaaccagggcagcctcccaaactcctgatctacagggcatccactctggcatctggggtcccatcgcggttcagaggcagtggatctgggacagagtacactctcaccatcaccgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactaatacttatggaaatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 582:

(SEQ ID NO: 592) gccaacatcgtgatgacccagactccatcccccgtgtctggagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtgactacttagcctggtatcagcagaaaccagggcagcctcccaaactcctgatctacagggcatccactctggcatctggggtcccatcgcggttcagaggcagtggatctgggacagagtacactctcaccatcaccgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactaatacttatggaaatactttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 590:

(SEQ ID NO: 600) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 574; SEQ ID NO: 576; and SEQ ID NO: 578, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 561or the variable heavy chain sequence of SEQ ID NO: 562, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 594; SEQ ID NO: 596;and SEQ ID NO: 598, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 581 or the variable light chain sequence of SEQ ID NO: 582,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 573; SEQ ID NO: 575; SEQ ID NO: 577; and SEQ ID NO: 579,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 561 orthe variable heavy chain sequence of SEQ ID NO: 562, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 593; SEQ ID NO: 595; SEQID NO: 597; and SEQ ID NO: 599, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 581 or the variable light chain sequence of SEQ ID NO: 582, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 571 encoding the heavy chainsequence of SEQ ID NO: 561; the polynucleotide SEQ ID NO: 572 encodingthe variable heavy chain sequence of SEQ ID NO: 562; the polynucleotideSEQ ID NO: 591 encoding the light chain sequence of SEQ ID NO: 581; thepolynucleotide SEQ ID NO: 592 encoding the variable light chain sequenceof SEQ ID NO: 582; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 574; SEQ ID NO: 576; andSEQ ID NO: 578) of the heavy chain sequence of SEQ ID NO: 561 or thevariable heavy chain sequence of SEQ ID NO: 562; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 594; SEQ IDNO: 596; and SEQ ID NO: 598) of the light chain sequence of SEQ ID NO:581 or the variable light chain sequence of SEQ ID NO: 582;polynucleotides encoding the framework regions (SEQ ID NO: 573; SEQ IDNO: 575; SEQ ID NO: 577; and SEQ ID NO: 579) of the heavy chain sequenceof SEQ ID NO: 561 or the variable heavy chain sequence of SEQ ID NO:562; and polynucleotides encoding the framework regions (SEQ ID NO: 593;SEQ ID NO: 595; SEQ ID NO: 597; and SEQ ID NO: 599) of the light chainsequence of SEQ ID NO: 581 or the variable light chain sequence of SEQID NO: 582.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab15, thepolynucleotides encoding the full length Ab15 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 571 encoding theheavy chain sequence of SEQ ID NO: 561 and the polynucleotide SEQ ID NO:591 encoding the light chain sequence of SEQ ID NO: 581.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab15 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab15 or Fab fragmentsthereof may be produced via expression of Ab15 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab16

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 601:

(SEQ ID NO: 611) caggagcagctggtggagtccgggggaggcctggtccagcctgagggatccctgacactcacctgcacagcctctggattctcctttagtagtgattactggatatgctgggtccgccaggctccagggaagggcctggagtggatcggatgcattcgtgatggtggtgggagttactacgcgaactgggcgaaaggccgactcaccatctccatgacctcgtcgaccacggtgggtctgaaaatgaccagtctgacagccgcggacacggccacgtatttttgtgcgagcgatattaatgatgggtggcttggccaattcaacttgtggggcccagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 602:

(SEQ ID NO: 612) caggagcagctggtggagtccgggggaggcctggtccagcctgagggatccctgacactcacctgcacagcctctggattctcctttagtagtgattactggatatgctgggtccgccaggctccagggaagggcctggagtggatcggatgcattcgtgatggtggtgggagttactacgcgaactgggcgaaaggccgactcaccatctccatgacctcgtcgaccacggtgggtctgaaaatgaccagtctgacagccgcggacacggccacgtatttttgtgcgagcgatattaatgatgggtggcttggccaattcaacttgtggggcccagggaccc tcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 610:

(SEQ ID NO: 620) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:621:

(SEQ ID NO: 631) gctgacattgtgatgacccagactccagcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtagctacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacagggcatccactctggcctctggggtcccatcgcggttcagcggcagtggatctgggacagagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactgttacttatggaaatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 622:

(SEQ ID NO: 632) gctgacattgtgatgacccagactccagcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtcagagcattagtagctacttagcctggtatcagcagaaaccagggcagcctcccaagctcctgatctacagggcatccactctggcctctggggtcccatcgcggttcagcggcagtggatctgggacagagttcactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactgttacttatggaaatactttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 630:

(SEQ ID NO: 640) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 614; SEQ ID NO: 616; and SEQ ID NO: 618, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 601or the variable heavy chain sequence of SEQ ID NO: 602, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 634; SEQ ID NO: 636;and SEQ ID NO: 638, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 621 or the variable light chain sequence of SEQ ID NO: 622,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 613; SEQ ID NO: 615; SEQ ID NO: 617; and SEQ ID NO: 619,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 601 orthe variable heavy chain sequence of SEQ ID NO: 602, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 633; SEQ ID NO: 635; SEQID NO: 637; and SEQ ID NO: 639, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 621 or the variable light chain sequence of SEQ ID NO: 622, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 611 encoding the heavy chainsequence of SEQ ID NO: 601; the polynucleotide SEQ ID NO: 612 encodingthe variable heavy chain sequence of SEQ ID NO: 602; the polynucleotideSEQ ID NO: 631 encoding the light chain sequence of SEQ ID NO: 621; thepolynucleotide SEQ ID NO: 632 encoding the variable light chain sequenceof SEQ ID NO: 622; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 614; SEQ ID NO: 616; andSEQ ID NO: 618) of the heavy chain sequence of SEQ ID NO: 601 or thevariable heavy chain sequence of SEQ ID NO: 602; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 634; SEQ IDNO: 636; and SEQ ID NO: 638) of the light chain sequence of SEQ ID NO:621 or the variable light chain sequence of SEQ ID NO: 622;polynucleotides encoding the framework regions (SEQ ID NO: 613; SEQ IDNO: 615; SEQ ID NO: 617; and SEQ ID NO: 619) of the heavy chain sequenceof SEQ ID NO: 601 or the variable heavy chain sequence of SEQ ID NO:602; and polynucleotides encoding the framework regions (SEQ ID NO: 633;SEQ ID NO: 635; SEQ ID NO: 637; and SEQ ID NO: 639) of the light chainsequence of SEQ ID NO: 621 or the variable light chain sequence of SEQID NO: 622.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab16, thepolynucleotides encoding the full length Ab16 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 611 encoding theheavy chain sequence of SEQ ID NO: 601 and the polynucleotide SEQ ID NO:631 encoding the light chain sequence of SEQ ID NO: 621.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab16 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab16 or Fab fragmentsthereof may be produced via expression of Ab16 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab17

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 641:

(SEQ ID NO: 651) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacagcctctggattctccttcagtagcagctactggatatgctgggtccgccaggctccagggaagggactggagtggatcggatgcattcgtcctggtagtgcggattactacgcgagctgggcgaaaggccgattcaccatctccagagcctcgtcgtccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttttgtgcgagcgatattaatgatgggtggcttggccaattcaacttgtggggcccaggcaccctggtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 642:

(SEQ ID NO: 652) caggagcagctggaggagtccgggggagacctggtcaagcctgagggatccctgacactcacctgcacagcctctggattctccttcagtagcagctactggatatgctgggtccgccaggctccagggaagggactggagtggatcggatgcattcgtcctggtagtgcggattactacgcgagctgggcgaaaggccgattcaccatctccagagcctcgtcgtccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttttgtgcgagcgatattaatgatgggtggcttggccaattcaacttgtggggcccaggcaccc tggtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 650:

(SEQ ID NO: 660) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:661:

(SEQ ID NO: 671) gccgatgttgtgatgacccagactccagcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtctgagcattgctgactacttagcctggtatctccagaaaccagggcagcctcccaagctcctgatctacagggcatccactctggcatctggggtcccatcgcggttcaagggcagtggatctgggacagagtacactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactaatacttatggaaatactttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacag gggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 662:

(SEQ ID NO: 672) gccgatgttgtgatgacccagactccagcctccgtggaggcagctgtgggaggcacagtcaccatcaagtgccaggccagtctgagcattgctgactacttagcctggtatctccagaaaccagggcagcctcccaagctcctgatctacagggcatccactctggcatctggggtcccatcgcggttcaagggcagtggatctgggacagagtacactctcaccatcagcgacctggagtgtgccgatgctgccacttactactgtcaaagctattattccgttactactaatacttatggaaatactttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 670:

(SEQ ID NO: 680) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 654; SEQ ID NO: 656; and SEQ ID NO: 658, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 641or the variable heavy chain sequence of SEQ ID NO: 642, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 674; SEQ ID NO: 676;and SEQ ID NO: 678, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 661 or the variable light chain sequence of SEQ ID NO: 662,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 653; SEQ ID NO: 655; SEQ ID NO: 657; and SEQ ID NO: 659,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 641 orthe variable heavy chain sequence of SEQ ID NO: 642, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 673; SEQ ID NO: 675; SEQID NO: 677; and SEQ ID NO: 679, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 661 or the variable light chain sequence of SEQ ID NO: 662, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 651 encoding the heavy chainsequence of SEQ ID NO: 641; the polynucleotide SEQ ID NO: 652 encodingthe variable heavy chain sequence of SEQ ID NO: 642; the polynucleotideSEQ ID NO: 671 encoding the light chain sequence of SEQ ID NO: 661; thepolynucleotide SEQ ID NO: 672 encoding the variable light chain sequenceof SEQ ID NO: 662; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 654; SEQ ID NO: 656; andSEQ ID NO: 658) of the heavy chain sequence of SEQ ID NO: 641 or thevariable heavy chain sequence of SEQ ID NO: 642; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 674; SEQ IDNO: 676; and SEQ ID NO: 678) of the light chain sequence of SEQ ID NO:661 or the variable light chain sequence of SEQ ID NO: 662;polynucleotides encoding the framework regions (SEQ ID NO: 653; SEQ IDNO: 655; SEQ ID NO: 657; and SEQ ID NO: 659) of the heavy chain sequenceof SEQ ID NO: 641 or the variable heavy chain sequence of SEQ ID NO:642; and polynucleotides encoding the framework regions (SEQ ID NO: 673;SEQ ID NO: 675; SEQ ID NO: 677; and SEQ ID NO: 679) of the light chainsequence of SEQ ID NO: 661 or the variable light chain sequence of SEQID NO: 662.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab17, thepolynucleotides encoding the full length Ab17 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 651 encoding theheavy chain sequence of SEQ ID NO: 641 and the polynucleotide SEQ ID NO:671 encoding the light chain sequence of SEQ ID NO: 661.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab17 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab17 or Fab fragmentsthereof may be produced via expression of Ab17 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab18

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 681:

(SEQ ID NO: 691) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaactactggatatgctgggtccgtcaggctccagggaaggggctggagtggatcggatgcattcgtgatggtggtggcacttactacgctagctctgctaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagcgatatcaatgatgggtggcttggccaattcaacttgtggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacgcgagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggt aaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 682:gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattcaccgtcagtagcaactactggatatgctgggtccgtcaggctccagggaaggggctggagtggatcggatgcattcgtgatggtggtggcacttactacgctagctctgctaaaggccgattcaccatctccagagacaattccaagaacaccctgtatcttcaaatgaacagcctgagagctgaggacactgctgtgtattactgtgctagcgatatcaatgatgggtggcttggccaattcaacttgtggggccaagggaccctcgtcaccgtctcgagc (SEQ ID NO: 692).

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 690:

(SEQ ID NO: 700) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacgcgagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:701:

(SEQ ID NO: 711) gctgacattgtgatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcaagtgccaggccagtcagagcattagtgcttacttagcctggtatcagcagaaaccagggaaagtccctaagctcctgatctatagggcatacactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaaagctactattccgttactactaatacttatggaaatactttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacag gggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 702:

(SEQ ID NO: 712) gctgacattgtgatgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcaagtgccaggccagtcagagcattagtgcttacttagcctggtatcagcagaaaccagggaaagtccctaagctcctgatctatagggcatacactctggcatctggggtcccatctcgtttcagtggcagtggatctgggacagatttcactctcaccatcagcagcctgcagcctgaagatgttgcaacttattactgtcaaagctactattccgttactactaatacttatggaaatactttcggcggaggaaccaaggtggaaatcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 710:

(SEQ ID NO: 720) cgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 694; SEQ ID NO: 696; and SEQ ID NO: 698, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 681or the variable heavy chain sequence of SEQ ID NO: 682, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 714; SEQ ID NO: 716;and SEQ ID NO: 718, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 701 or the variable light chain sequence of SEQ ID NO: 702,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 693; SEQ ID NO: 695; SEQ ID NO: 697; and SEQ ID NO: 699,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 681 orthe variable heavy chain sequence of SEQ ID NO: 682, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 713; SEQ ID NO: 715; SEQID NO: 717; and SEQ ID NO: 719, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 701 or the variable light chain sequence of SEQ ID NO: 702, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 691 encoding the heavy chainsequence of SEQ ID NO: 681; the polynucleotide SEQ ID NO: 692 encodingthe variable heavy chain sequence of SEQ ID NO: 682; the polynucleotideSEQ ID NO: 711 encoding the light chain sequence of SEQ ID NO: 701; thepolynucleotide SEQ ID NO: 712 encoding the variable light chain sequenceof SEQ ID NO: 702; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 694; SEQ ID NO: 696; andSEQ ID NO: 698) of the heavy chain sequence of SEQ ID NO: 681 or thevariable heavy chain sequence of SEQ ID NO: 682; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 714; SEQ IDNO: 716; and SEQ ID NO: 718) of the light chain sequence of SEQ ID NO:701 or the variable light chain sequence of SEQ ID NO: 702;polynucleotides encoding the framework regions (SEQ ID NO: 693; SEQ IDNO: 695; SEQ ID NO: 697; and SEQ ID NO: 699) of the heavy chain sequenceof SEQ ID NO: 681 or the variable heavy chain sequence of SEQ ID NO:682; and polynucleotides encoding the framework regions (SEQ ID NO: 713;SEQ ID NO: 715; SEQ ID NO: 717; and SEQ ID NO: 719) of the light chainsequence of SEQ ID NO: 701 or the variable light chain sequence of SEQID NO: 702.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab18, thepolynucleotides encoding the full length Ab18 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 691 encoding theheavy chain sequence of SEQ ID NO: 681 and the polynucleotide SEQ ID NO:711 encoding the light chain sequence of SEQ ID NO: 701.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab18 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab18 or Fab fragmentsthereof may be produced via expression of Ab18 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab19

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 721:

(SEQ ID NO: 731) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggaatcgacctcagtagctatgcaatgggctgggtccgtcaggctccagggaaggggctggagtacatcggaatcattgttagttatgggcccacatactacgctagctgggctaaaggccgattcaccatctccagagacaattccaagaacaccgtgtatcttcaaatgaacagcctgagagctgaggacactgccacctatttctgtgccagagatctggatgctcaaagtagtggttactatggagcttttaacatctggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtct ccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 722:

(SEQ ID NO: 732) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggaatcgacctcagtagctatgcaatgggctgggtccgtcaggctccagggaaggggctggagtacatcggaatcattgttagttatgggcccacatactacgctagctgggctaaaggccgattcaccatctccagagacaattccaagaacaccgtgtatcttcaaatgaacagcctgagagctgaggacactgccacctatttctgtgccagagatctggatgctcaaagtagtggttactatggagcttttaacatctggggccaagggac cctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 730:

(SEQ ID NO: 740) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:741:

(SEQ ID NO: 751) gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgtcaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatgctgcatcccctctggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttactactgtcaaagctattatggtagtagcaacattgctttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 742:

(SEQ ID NO: 752) gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgtcaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatgctgcatcccctctggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttactactgtcaaagctattatggtagtagcaacattgctttcggcggaggaaccaaggtggaaatcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 750:

(SEQ ID NO: 760) cgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 734; SEQ ID NO: 736; and SEQ ID NO: 738, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 721or the variable heavy chain sequence of SEQ ID NO: 722, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 754; SEQ ID NO: 756;and SEQ ID NO: 758, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 741 or the variable light chain sequence of SEQ ID NO: 742,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 733; SEQ ID NO: 735; SEQ ID NO: 737; and SEQ ID NO: 739,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 721 orthe variable heavy chain sequence of SEQ ID NO: 722, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 753; SEQ ID NO: 755; SEQID NO: 757; and SEQ ID NO: 759, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 741 or the variable light chain sequence of SEQ ID NO: 742, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 731 encoding the heavy chainsequence of SEQ ID NO: 721; the polynucleotide SEQ ID NO: 732 encodingthe variable heavy chain sequence of SEQ ID NO: 722; the polynucleotideSEQ ID NO: 751 encoding the light chain sequence of SEQ ID NO: 741; thepolynucleotide SEQ ID NO: 752 encoding the variable light chain sequenceof SEQ ID NO: 742; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 734; SEQ ID NO: 736; andSEQ ID NO: 738) of the heavy chain sequence of SEQ ID NO: 721 or thevariable heavy chain sequence of SEQ ID NO: 722; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 754; SEQ IDNO: 756; and SEQ ID NO: 758) of the light chain sequence of SEQ ID NO:741 or the variable light chain sequence of SEQ ID NO: 742;polynucleotides encoding the framework regions (SEQ ID NO: 733; SEQ IDNO: 735; SEQ ID NO: 737; and SEQ ID NO: 739) of the heavy chain sequenceof SEQ ID NO: 721 or the variable heavy chain sequence of SEQ ID NO:722; and polynucleotides encoding the framework regions (SEQ ID NO: 753;SEQ ID NO: 755; SEQ ID NO: 757; and SEQ ID NO: 759) of the light chainsequence of SEQ ID NO: 741 or the variable light chain sequence of SEQID NO: 742.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab19, thepolynucleotides encoding the full length Ab19 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 731 encoding theheavy chain sequence of SEQ ID NO: 721 and the polynucleotide SEQ ID NO:751 encoding the light chain sequence of SEQ ID NO: 741.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab19 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab19 or Fab fragmentsthereof may be produced via expression of Ab19 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab20

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 761:

(SEQ ID NO: 771) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggaatcgacctcagtagctatgcaatgggctgggtccgtcaggctccagggaaggggctggagtacatcggaatcattgttagttatgggcccacatactacgctagctgggctaaaggccgattcaccatctccagagacaattccaagtccaccgtgtatcttcaaatgaacagcctgagagctgaggacactgccacctatttctgtgccagagatctggatgctcaaagtagtggttactatggagcttttaacatctggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtct ccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 762:

(SEQ ID NO: 772) gaggtgcagcttgtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggaatcgacctcagtagctatgcaatgggctgggtccgtcaggctccagggaaggggctggagtacatcggaatcattgttagttatgggcccacatactacgctagctgggctaaaggccgattcaccatctccagagacaattccaagtccaccgtgtatcttcaaatgaacagcctgagagctgaggacactgccacctatttctgtgccagagatctggatgctcaaagtagtggttactatggagcttttaacatctggggccaagggaccctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 770:

(SEQ ID NO: 780) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:781:

(SEQ ID NO: 791) gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgtcaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatgctgcatcccctctggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttactactgtcaaagctattatggtagtagcaacattgctttcggcggaggaaccaaggtggaaatcaaacgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggg gagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 782:

(SEQ ID NO: 792) gacatccagatgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgtcaggccagtcagagcattagcactgcattagcctggtatcagcagaaaccaggaaaagcccctaagctcctgatctatgctgcatcccctctggcatctggagtcccatcaaggttcagcggcagtggatctggaacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttactactgtcaaagctattatggtagtagcaacattgctttcggcggaggaaccaaggtggaaatcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 790:

(SEQ ID NO: 800) cgtacggtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 774; SEQ ID NO: 776; and SEQ ID NO: 778, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 761or the variable heavy chain sequence of SEQ ID NO: 762, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 794; SEQ ID NO: 796;and SEQ ID NO: 798, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 781 or the variable light chain sequence of SEQ ID NO: 782,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 773; SEQ ID NO: 775; SEQ ID NO: 777; and SEQ ID NO: 779,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 761 orthe variable heavy chain sequence of SEQ ID NO: 762, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 793; SEQ ID NO: 795; SEQID NO: 797; and SEQ ID NO: 799, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 781 or the variable light chain sequence of SEQ ID NO: 782, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 771 encoding the heavy chainsequence of SEQ ID NO: 761; the polynucleotide SEQ ID NO: 772 encodingthe variable heavy chain sequence of SEQ ID NO: 762; the polynucleotideSEQ ID NO: 791 encoding the light chain sequence of SEQ ID NO: 781; thepolynucleotide SEQ ID NO: 792 encoding the variable light chain sequenceof SEQ ID NO: 782; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 774; SEQ ID NO: 776; andSEQ ID NO: 778) of the heavy chain sequence of SEQ ID NO: 761 or thevariable heavy chain sequence of SEQ ID NO: 762; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 794; SEQ IDNO: 796; and SEQ ID NO: 798) of the light chain sequence of SEQ ID NO:781 or the variable light chain sequence of SEQ ID NO: 782;polynucleotides encoding the framework regions (SEQ ID NO: 773; SEQ IDNO: 775; SEQ ID NO: 777; and SEQ ID NO: 779) of the heavy chain sequenceof SEQ ID NO: 761 or the variable heavy chain sequence of SEQ ID NO:762; and polynucleotides encoding the framework regions (SEQ ID NO: 793;SEQ ID NO: 795; SEQ ID NO: 797; and SEQ ID NO: 799) of the light chainsequence of SEQ ID NO: 781 or the variable light chain sequence of SEQID NO: 782.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab20, thepolynucleotides encoding the full length Ab20 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 771 encoding theheavy chain sequence of SEQ ID NO: 761 and the polynucleotide SEQ ID NO:791 encoding the light chain sequence of SEQ ID NO: 781.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab20 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab20 or Fab fragmentsthereof may be produced via expression of Ab20 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab21

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 801:

(SEQ ID NO: 811) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtagttatgcaatgaattgggtccgccaggctccagggaaggggctggagtggatcggggccattcgtagtagtggtgccacattcttcgcgagctgggtgaatggccgtttcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacacccgaggacacggccacctatttctgtgccagagatactaatgatggttggtatattaatcggttggatctctggggcccgggcaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 802:

(SEQ ID NO: 812) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacctgcacagtctctggattctccctcagtagttatgcaatgaattgggtccgccaggctccagggaaggggctggagtggatcggggccattcgtagtagtggtgccacattcttcgcgagctgggtgaatggccgtttcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtccgacacccgaggacacggccacctatttctgtgccagagatactaatgatggttggtatattaatcggttggatctctggggcccgggcaccctcgtca ccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 810:

(SEQ ID NO: 820) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtc tccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:821:

(SEQ ID NO: 831) gccgccgtgctgacccagactccatctcccgtgtctgcagctgtgggaggcacagtcagcatcagttgccagtccagtaagagtgtttatagtaactacttatcctggtttcagcagaaaccagggcagcctcccaagttcctgatctacaaggcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacgtgcagtgtgacgatgctgccacttactactgtgcaggcggtgatactaatattagtgataatgctttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttca acaggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 822:

(SEQ ID NO: 832) gccgccgtgctgacccagactccatctcccgtgtctgcagctgtgggaggcacagtcagcatcagttgccagtccagtaagagtgtttatagtaactacttatcctggtttcagcagaaaccagggcagcctcccaagttcctgatctacaaggcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacgtgcagtgtgacgatgctgccacttactactgtgcaggcggtgatactaatattagtgataatgctttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 830:

(SEQ ID NO: 840) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 814; SEQ ID NO: 816; and SEQ ID NO: 818, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 801or the variable heavy chain sequence of SEQ ID NO: 802, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 834; SEQ ID NO: 836;and SEQ ID NO: 838, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 821 or the variable light chain sequence of SEQ ID NO: 822,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 813; SEQ ID NO: 815; SEQ ID NO: 817; and SEQ ID NO: 819,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 801 orthe variable heavy chain sequence of SEQ ID NO: 802, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 833; SEQ ID NO: 835; SEQID NO: 837; and SEQ ID NO: 839, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 821 or the variable light chain sequence of SEQ ID NO: 822, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 811 encoding the heavy chainsequence of SEQ ID NO: 801; the polynucleotide SEQ ID NO: 812 encodingthe variable heavy chain sequence of SEQ ID NO: 802; the polynucleotideSEQ ID NO: 831 encoding the light chain sequence of SEQ ID NO: 821; thepolynucleotide SEQ ID NO: 832 encoding the variable light chain sequenceof SEQ ID NO: 822; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 814; SEQ ID NO: 816; andSEQ ID NO: 818) of the heavy chain sequence of SEQ ID NO: 801 or thevariable heavy chain sequence of SEQ ID NO: 802; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 834; SEQ IDNO: 836; and SEQ ID NO: 838) of the light chain sequence of SEQ ID NO:821 or the variable light chain sequence of SEQ ID NO: 822;polynucleotides encoding the framework regions (SEQ ID NO: 813; SEQ IDNO: 815; SEQ ID NO: 817; and SEQ ID NO: 819) of the heavy chain sequenceof SEQ ID NO: 801 or the variable heavy chain sequence of SEQ ID NO:802; and polynucleotides encoding the framework regions (SEQ ID NO: 833;SEQ ID NO: 835; SEQ ID NO: 837; and SEQ ID NO: 839) of the light chainsequence of SEQ ID NO: 821 or the variable light chain sequence of SEQID NO: 822.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab21, thepolynucleotides encoding the full length Ab21 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 811 encoding theheavy chain sequence of SEQ ID NO: 801 and the polynucleotide SEQ ID NO:831 encoding the light chain sequence of SEQ ID NO: 821.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab21 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab21 or Fab fragmentsthereof may be produced via expression of Ab21 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab22

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 841:

(SEQ ID NO: 851) gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattctccctcagtagttatgcaatgaattgggtccgccaggctccagggaaggggctggagtggatcggggccattcgtagtagtggtgccacattcttcgcgagctccgtgaatggcagattcaccatctccagagacaattccaagaacacggtgtatcttcaaatgaacagcctgagagccgaggacacggctgtgtattactgtgcgagagatactaatgatggttggtatattaatcggttggatctctggggccaagggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacgcgagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 842:

(SEQ ID NO: 852) gaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctgagactctcctgtgcagcctctggattctccctcagtagttatgcaatgaattgggtccgccaggctccagggaaggggctggagtggatcggggccattcgtagtagtggtgccacattcttcgcgagctccgtgaatggcagattcaccatctccagagacaattccaagaacacggtgtatcttcaaatgaacagcctgagagccgaggacacggctgtgtattactgtgcgagagatactaatgatggttggtatattaatcggttggatctctggggccaaggga ccctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 850:

(SEQ ID NO: 860) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacgcgagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgt ctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:861:

(SEQ ID NO: 871) gccgtgctgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccagtccagtaagagtgtttatagtaactacttatcctggtttcagcagaaaccagggaaagcccctaagttcctgatctataaggcatccactctggcatctggggtcccatcaaggttcagcggcagtggatctgggacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgcgcaggcggtgatactaatattgctgataatgctttcggcggaggaaccaaggtggaaatcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaa caggggagagtgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 862:

(SEQ ID NO: 872) gccgtgctgacccagtctccttccaccctgtctgcatctgtaggagacagagtcaccatcacttgccagtccagtaagagtgtttatagtaactacttatcctggtttcagcagaaaccagggaaagcccctaagttcctgatctataaggcatccactctggcatctggggtcccatcaaggttcagcggcagtggatctgggacagaattcactctcaccatcagcagcctgcagcctgatgattttgcaacttattactgcgcaggcggtgatactaatattgctgataatgctttcggcggaggaaccaaggtggaaatcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 870:

(SEQ ID NO: 880) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 854; SEQ ID NO: 856; and SEQ ID NO: 858, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 841or the variable heavy chain sequence of SEQ ID NO: 842, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 874; SEQ ID NO: 876;and SEQ ID NO: 878, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 861 or the variable light chain sequence of SEQ ID NO: 862,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 853; SEQ ID NO: 855; SEQ ID NO: 857; and SEQ ID NO: 859,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 841 orthe variable heavy chain sequence of SEQ ID NO: 842, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 873; SEQ ID NO: 875; SEQID NO: 877; and SEQ ID NO: 879, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 861 or the variable light chain sequence of SEQ ID NO: 862, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 851 encoding the heavy chainsequence of SEQ ID NO: 841; the polynucleotide SEQ ID NO: 852 encodingthe variable heavy chain sequence of SEQ ID NO: 842; the polynucleotideSEQ ID NO: 871 encoding the light chain sequence of SEQ ID NO: 861; thepolynucleotide SEQ ID NO: 872 encoding the variable light chain sequenceof SEQ ID NO: 862; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 854; SEQ ID NO: 856; andSEQ ID NO: 858) of the heavy chain sequence of SEQ ID NO: 841 or thevariable heavy chain sequence of SEQ ID NO: 842; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 874; SEQ IDNO: 876; and SEQ ID NO: 878) of the light chain sequence of SEQ ID NO:861 or the variable light chain sequence of SEQ ID NO: 862;polynucleotides encoding the framework regions (SEQ ID NO: 853; SEQ IDNO: 855; SEQ ID NO: 857; and SEQ ID NO: 859) of the heavy chain sequenceof SEQ ID NO: 841 or the variable heavy chain sequence of SEQ ID NO:842; and polynucleotides encoding the framework regions (SEQ ID NO: 873;SEQ ID NO: 875; SEQ ID NO: 877; and SEQ ID NO: 879) of the light chainsequence of SEQ ID NO: 861 or the variable light chain sequence of SEQID NO: 862.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab22, thepolynucleotides encoding the full length Ab22 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 851 encoding theheavy chain sequence of SEQ ID NO: 841 and the polynucleotide SEQ ID NO:871 encoding the light chain sequence of SEQ ID NO: 861.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab22 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab22 or Fab fragmentsthereof may be produced via expression of Ab22 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab23

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 881:

(SEQ ID NO: 891) cagtcgttggaggagtccgggggagacctggtcaagcctggggcatccctgacactcacctgcaaagcctctggattctccttcagtagcggctactacatgtgctgggtccgccaggctccagggaaggggctggagtggatcgcatgcatttatgctggtagtggtggtagcactttcttcgcgaactgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttctgtgcgagagatggtggttatgctggctatggttatgctttctttaacttgtggggcccggggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtct ccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 882:

(SEQ ID NO: 892) cagtcgttggaggagtccgggggagacctggtcaagcctggggcatccctgacactcacctgcaaagcctctggattctccttcagtagcggctactacatgtgctgggtccgccaggctccagggaaggggctggagtggatcgcatgcatttatgctggtagtggtggtagcactttcttcgcgaactgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttctgtgcgagagatggtggttatgctggctatggttatgctttctttaacttgtggggcccggggac cctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 890:

(SEQ ID NO: 900) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:901:

(SEQ ID NO: 911) gatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtgagaggatttatagtggtttggcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagatttcactctcaccatcagcgacctggagtgtgacgatgctgccatttactactgtcaatgtacttattatggttctagttatcctaatgttttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagag tgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 902:

(SEQ ID NO: 912) gatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtgagaggatttatagtggtttggcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagatttcactctcaccatcagcgacctggagtgtgacgatgctgccatttactactgtcaatgtacttattatggttctagttatcctaatgttttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 910:

(SEQ ID NO: 920) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 894; SEQ ID NO: 896; and SEQ ID NO: 898, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 881or the variable heavy chain sequence of SEQ ID NO: 882, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 914; SEQ ID NO: 916;and SEQ ID NO: 918, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 901 or the variable light chain sequence of SEQ ID NO: 902,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 893; SEQ ID NO: 895; SEQ ID NO: 897; and SEQ ID NO: 899,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 881 orthe variable heavy chain sequence of SEQ ID NO: 882, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 913; SEQ ID NO: 915; SEQID NO: 917; and SEQ ID NO: 919, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 901 or the variable light chain sequence of SEQ ID NO: 902, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 891 encoding the heavy chainsequence of SEQ ID NO: 881; the polynucleotide SEQ ID NO: 892 encodingthe variable heavy chain sequence of SEQ ID NO: 882; the polynucleotideSEQ ID NO: 911 encoding the light chain sequence of SEQ ID NO: 901; thepolynucleotide SEQ ID NO: 912 encoding the variable light chain sequenceof SEQ ID NO: 902; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 894; SEQ ID NO: 896; andSEQ ID NO: 898) of the heavy chain sequence of SEQ ID NO: 881 or thevariable heavy chain sequence of SEQ ID NO: 882; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 914; SEQ IDNO: 916; and SEQ ID NO: 918) of the light chain sequence of SEQ ID NO:901 or the variable light chain sequence of SEQ ID NO: 902;polynucleotides encoding the framework regions (SEQ ID NO: 893; SEQ IDNO: 895; SEQ ID NO: 897; and SEQ ID NO: 899) of the heavy chain sequenceof SEQ ID NO: 881 or the variable heavy chain sequence of SEQ ID NO:882; and polynucleotides encoding the framework regions (SEQ ID NO: 913;SEQ ID NO: 915; SEQ ID NO: 917; and SEQ ID NO: 919) of the light chainsequence of SEQ ID NO: 901 or the variable light chain sequence of SEQID NO: 902.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab23, thepolynucleotides encoding the full length Ab23 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 891 encoding theheavy chain sequence of SEQ ID NO: 881 and the polynucleotide SEQ ID NO:911 encoding the light chain sequence of SEQ ID NO: 901.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab23 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab23 or Fab fragmentsthereof may be produced via expression of Ab23 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab24

In one embodiment, the invention is further directed to polynucleotidesencoding antibody polypeptides having binding specificity to PCSK9. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain sequence of SEQ ID NO: 921:

(SEQ ID NO: 931) cagtcgttggaggagtccgggggagacctggtcaagcctggggcatccctgacactcacctgcaaagcctctggattctccttcagtagcggctactacatgtgctgggtccgccaggctccagggaaggggctggagtggatcgcatgcatttatgctggtagtggtggtagcactttcttcgcgaactgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttctgtgcgagagatggtggttatgctggctatggttatgctttctttaacttgtggggcccggggaccctcgtcaccgtctcgagcgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtct ccgggtaaa.

In another embodiment of the invention, the polynucleotides of theinvention comprise, or alternatively consist of, the followingpolynucleotide sequence encoding the variable heavy chain polypeptidesequence of SEQ ID NO: 922:

(SEQ ID NO: 932) cagtcgttggaggagtccgggggagacctggtcaagcctggggcatccctgacactcacctgcaaagcctctggattctccttcagtagcggctactacatgtgctgggtccgccaggctccagggaaggggctggagtggatcgcatgcatttatgctggtagtggtggtagcactttcttcgcgaactgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtgactctgcaaatgaccagtctgacagccgcggacacggccacctatttctgtgcgagagatggtggttatgctggctatggttatgctttctttaacttgtggggcccggggac cctcgtcaccgtctcgagc.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant heavy chain polypeptide sequence of SEQID NO: 930:

(SEQ ID NO: 940) gcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacgccagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:941:

(SEQ ID NO: 951) gatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtgagaggatttatagtggtttggcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagatttcactctcaccatcagcgacctggagtgtgacgatgctgccatttactactgtcaagctacttattatggttctagttatcctaatgttttcggcggagggaccgaggtggtggtcaaacgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagag tgt.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 942:

(SEQ ID NO: 952) gatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagtcaccatcaagtgccaggccagtgagaggatttatagtggtttggcctggtatcagcagaaaccagggcagcctcccaagctcctgatctatggtgcatccactctggcatctggggtcccatcgcggttcaaaggcagtggatctgggacagatttcactctcaccatcagcgacctggagtgtgacgatgctgccatttactactgtcaagctacttattatggttctagttatcctaatgttttcggcggagggaccgaggtggtggtcaaa.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the constant light chain polypeptide sequence of SEQID NO: 950:

(SEQ ID NO: 960) cgtacggtagcggccccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 934; SEQ ID NO: 936; and SEQ ID NO: 938, which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain sequence of SEQ ID NO: 921or the variable heavy chain sequence of SEQ ID NO: 922, and/or one ormore of the polynucleotide sequences of SEQ ID NO: 954; SEQ ID NO: 956;and SEQ ID NO: 958, which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the light chain sequence ofSEQ ID NO: 941 or the variable light chain sequence of SEQ ID NO: 942,or combinations of these polynucleotide sequences. In another embodimentof the invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of polynucleotides encoding one or more of the CDRs, thevariable heavy chain and variable light chain sequences, and the heavychain and light chain sequences set forth above, including all of them.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to PCSK9 comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 933; SEQ ID NO: 935; SEQ ID NO: 937; and SEQ ID NO: 939,which correspond to polynucleotides encoding the framework regions (FRsor constant regions) of the heavy chain sequence of SEQ ID NO: 921 orthe variable heavy chain sequence of SEQ ID NO: 922, and/or one or moreof the polynucleotide sequences of SEQ ID NO: 953; SEQ ID NO: 955; SEQID NO: 957; and SEQ ID NO: 959, which correspond to the frameworkregions (FRs or constant regions) of the light chain sequence of SEQ IDNO: 941 or the variable light chain sequence of SEQ ID NO: 942, orcombinations of these polynucleotide sequences. In another embodiment ofthe invention, the polynucleotides encoding the antibodies of theinvention or fragments thereof comprise, or alternatively consist of,combinations of one or more of the FRs, the variable heavy chain andvariable light chain sequences, and the heavy chain and light chainsequences set forth above, including all of them.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to PCSK9comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 931 encoding the heavy chainsequence of SEQ ID NO: 921; the polynucleotide SEQ ID NO: 932 encodingthe variable heavy chain sequence of SEQ ID NO: 922; the polynucleotideSEQ ID NO: 951 encoding the light chain sequence of SEQ ID NO: 941; thepolynucleotide SEQ ID NO: 952 encoding the variable light chain sequenceof SEQ ID NO: 942; polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 934; SEQ ID NO: 936; andSEQ ID NO: 938) of the heavy chain sequence of SEQ ID NO: 921 or thevariable heavy chain sequence of SEQ ID NO: 922; polynucleotidesencoding the complementarity-determining regions (SEQ ID NO: 954; SEQ IDNO: 956; and SEQ ID NO: 958) of the light chain sequence of SEQ ID NO:941 or the variable light chain sequence of SEQ ID NO: 942;polynucleotides encoding the framework regions (SEQ ID NO: 933; SEQ IDNO: 935; SEQ ID NO: 937; and SEQ ID NO: 939) of the heavy chain sequenceof SEQ ID NO: 921 or the variable heavy chain sequence of SEQ ID NO:922; and polynucleotides encoding the framework regions (SEQ ID NO: 953;SEQ ID NO: 955; SEQ ID NO: 957; and SEQ ID NO: 959) of the light chainsequence of SEQ ID NO: 941 or the variable light chain sequence of SEQID NO: 942.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for PCSK9. With respect to antibody Ab24, thepolynucleotides encoding the full length Ab24 antibody comprise, oralternatively consist of, the polynucleotide SEQ ID NO: 931 encoding theheavy chain sequence of SEQ ID NO: 921 and the polynucleotide SEQ ID NO:951 encoding the light chain sequence of SEQ ID NO: 941.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab24 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-PCSK9 antibodies such as Ab24 or Fab fragmentsthereof may be produced via expression of Ab24 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

In one embodiment, the invention is directed to an isolatedpolynucleotide comprising a polynucleotide encoding an anti-PCSK9 V_(H)antibody amino acid sequence selected from SEQ ID NO: 12, SEQ ID NO: 52,SEQ ID NO: 92, SEQ ID NO: 132, SEQ ID NO: 172, SEQ ID NO: 212, SEQ IDNO: 252, SEQ ID NO: 292, SEQ ID NO: 332, SEQ ID NO: 372, SEQ ID NO: 412,SEQ ID NO: 452, SEQ ID NO: 492, SEQ ID NO: 532, SEQ ID NO: 572, SEQ IDNO: 612, SEQ ID NO: 652, SEQ ID NO: 692, SEQ ID NO: 732, SEQ ID NO: 772,SEQ ID NO: 812, SEQ ID NO: 852, SEQ ID NO: 892, SEQ ID NO: 932, orencoding a variant thereof wherein at least one framework residue (FRresidue) has been substituted with an amino acid present at thecorresponding position in a rabbit anti-PCSK9 antibody V_(H) polypeptideor a conservative amino acid substitution.

In another embodiment, the invention is directed to an isolatedpolynucleotide comprising the polynucleotide sequence encoding ananti-PCSK9 V_(L) antibody amino acid selected from SEQ ID NO: 32, SEQ IDNO: 72, SEQ ID NO: 112, SEQ ID NO: 152, SEQ ID NO: 192, SEQ ID NO: 232,SEQ ID NO: 272, SEQ ID NO: 312, SEQ ID NO: 352, SEQ ID NO: 392, SEQ IDNO: 432, SEQ ID NO: 472, SEQ ID NO: 512, SEQ ID NO: 552, SEQ ID NO: 592,SEQ ID NO: 632, SEQ ID NO: 672, SEQ ID NO: 712, SEQ ID NO: 752, SEQ IDNO: 792, SEQ ID NO: 832, SEQ ID NO: 872, SEQ ID NO: 912, SEQ ID NO: 952,or encoding a variant thereof wherein at least one framework residue (FRresidue) has been substituted with an amino acid present at thecorresponding position in a rabbit anti-PCSK9 antibody V_(L) polypeptideor a conservative amino acid substitution.

In yet another embodiment, the invention is directed to one or moreheterologous polynucleotides comprising a sequence encoding thepolypeptides contained in SEQ ID NO: 2 and SEQ ID NO: 22; SEQ ID NO: 42and SEQ ID NO: 62; SEQ ID NO: 82 and SEQ ID NO: 102; SEQ ID NO: 122 andSEQ ID NO: 142; SEQ ID NO: 162 and SEQ ID NO: 182; SEQ ID NO: 202 andSEQ ID NO: 222, SEQ ID NO: 242 and SEQ ID NO: 262; SEQ ID NO: 282 andSEQ ID NO: 302; SEQ ID NO: 322 and SEQ ID NO: 342; SEQ ID NO: 362 andSEQ ID NO: 382; SEQ ID NO: 402 and SEQ ID NO: 422; SEQ ID NO: 442 andSEQ ID NO: 462; SEQ ID NO: 482 and SEQ ID NO: 502; SEQ ID NO: 522 andSEQ ID NO: 542; SEQ ID NO: 562 and SEQ ID NO: 582; SEQ ID NO: 602 andSEQ ID NO: 622; SEQ ID NO: 642 and SEQ ID NO: 662; SEQ ID NO: 682 andSEQ ID NO: 702; SEQ ID NO: 722 and SEQ ID NO: 742; SEQ ID NO: 762 andSEQ ID NO: 782; SEQ ID NO: 802 and SEQ ID NO: 822; SEQ ID NO: 842 andSEQ ID NO: 862; SEQ ID NO: 882 and SEQ ID NO: 902; or SEQ ID NO: 922 andSEQ ID NO: 942.

In another embodiment, the invention is directed to an isolatedpolynucleotide that expresses a polypeptide containing at least one CDRpolypeptide derived from an anti-PCSK9 antibody wherein said expressedpolypeptide alone specifically binds PCSK9 or specifically binds PCSK9when expressed in association with another polynucleotide sequence thatexpresses a polypeptide containing at least one CDR polypeptide derivedfrom an anti-PCSK9 antibody wherein said at least one CDR is selectedfrom those contained in the V_(L) or V_(H) polypeptides of SEQ ID NOS:2, 22, 42, 62, 82, 102, 122, 142, 162, 182, 202, 222, 242, 262, 282,302, 322, 342, 362, 382, 402, 422, 442, 462, 482, 502, 522, 542, 562,582, 602, 622, 642, 662, 682, 702, 722, 742, 762, 782, 802, 822, 842,862, 882, 902, 922, or 942. More specifically, the at least one CDRcomprises SEQ ID NOS: 4, 6, 8, 24, 26, 28, 44, 46, 48, 64, 66, 68, 84,86, 88, 104, 106, 108, 124, 126, 128, 144, 146, 148, 164, 166, 168, 184,186, 188, 204, 206, 208, 224, 226, 228, 244, 246, 248, 264, 266, 268,284, 286, 288, 304, 306, 308, 324, 326, 328, 344, 346, 348, 364, 366,368, 384, 386, 388, 404, 406, 408, 424, 426, 428, 444, 446, 448, 464,466, 468, 484, 486, 488, 504, 506, 508, 524, 526, 528, 544, 546, 548,564, 566, 568, 584, 586, 588, 604, 606, 608, 624, 626, 628, 644, 646,648, 664, 666, 668, 684, 686, 688, 704, 706, 708, 724, 726, 728, 744,746, 748, 764, 766, 768, 784, 786, 788, 804, 806, 808, 824, 826, 828,844, 846, 848, 864, 866, 884, 886, 888, 904, 906, 908, 924, 926, 928,944, 946, or 948.

Host cells and vectors comprising said polynucleotides are alsocontemplated.

The invention further contemplates vectors comprising the polynucleotidesequences encoding the variable heavy and light chain polypeptidesequences, as well as the individual complementarity-determining regions(CDRs, or hypervariable regions), as set forth herein, as well as hostcells comprising said vector sequences. In one embodiment of theinvention, the host cell is a yeast cell. In another embodiment of theinvention, the yeast host cell belongs to the genus Pichia.

B-Cell Screening and Isolation

In one embodiment, the present invention contemplates the preparationand isolation of a clonal population of antigen-specific B cells thatmay be used for isolating at least one PCSK9 antigen-specific cell,which can be used to produce a monoclonal antibody against PCSK9, whichis specific to a desired PCSK9 antigen, or a nucleic acid sequencecorresponding to such an antibody. Methods of preparing and isolatingsaid clonal population of antigen-specific B cells are taught, forexample, in U.S. patent publication no. US 2007/0269868 toCarvalho-Jensen et al., the disclosure of which is herein incorporatedby reference in its entirety. Methods of preparing and isolating saidclonal population of antigen-specific B cells are also taught herein inthe examples. Methods of “enriching” a cell population by size ordensity are known in the art. See, e.g., U.S. Pat. No. 5,627,052. Thesesteps can be used in addition to enriching the cell population byantigen-specificity.

Methods of Humanizing Antibodies

In another embodiment, the present invention contemplates methods forhumanizing antibody heavy and light chains. Methods for humanizingantibody heavy and light chains which may be applied to anti-PCSK9antibodies are taught, for example, in U.S. patent applicationpublication no. US 2009/0022659 to Olson et al., and in U.S. Pat. No.7,935,340 to Garcia-Martinez et al., the disclosures of each of whichare herein incorporated by reference in their entireties.

Methods of Producing Antibodies and Fragments Thereof

In another embodiment, the present invention contemplates methods forproducing anti-PCSK9 antibodies and fragments thereof. Methods forproducing anti-PCSK9 antibodies and fragments thereof secreted frompolyploidal, preferably diploid or tetraploid strains of matingcompetent yeast are taught, for example, in U.S. patent applicationpublication no. US 2009/0022659 to Olson et al., and in U.S. Pat. No.7,935,340 to Garcia-Martinez et al., the disclosures of each of whichare herein incorporated by reference in their entireties. A preferredyeast for manufacture of antibodies is Pichia, and more preferablyPichia pastoris. However, antibodies according to the inventionpotentially may be made in other yeast such as other mating competentyeast of the Saccharomycetaceae family, which includes the generaArxiozyma; Ascobotryozyma; Citeromyces; Debaryomyces; Dekkera;Eremothecium; Issatchenkia; Kazachstania; Kluyveromyces; Kodamaea;Lodderomyces; Pachysolen; Pichia; Saccharomyces; Saturnispora;Tetrapisispora; Torulaspora; Williopsis; and Zygosaccharomyces. Othertypes of yeast potentially useful for making antibody proteins accordingto the invention include Yarrowia; Rhodosporidium; Candida; Hansenula;Filobasium; Sporidiobolus; Bullera; Leucosporidium and Filobasidella.

Other methods of producing antibodies are well known to those ofordinary skill in the art. For example, methods of producing chimericantibodies are now well known in the art (See, for example, U.S. Pat.No. 4,816,567 to Cabilly et al.; Morrison et al., P.N.A.S. USA,81:8651-55 (1984); Neuberger, M. S. et al., Nature, 314:268-270 (1985);Boulianne, G. L. et al., Nature, 312:643-46 (1984), the disclosures ofeach of which are herein incorporated by reference in their entireties).

Likewise, other methods of producing humanized antibodies are now wellknown in the art (See, for example, U.S. Pat. Nos. 5,530,101, 5,585,089,5,693,762, and 6,180,370 to Queen et al; U.S. Pat. Nos. 5,225,539 and6,548,640 to Winter; U.S. Pat. Nos. 6,054,297, 6,407,213 and 6,639,055to Carter et al; U.S. Pat. No. 6,632,927 to Adair; Jones, P. T. et al,Nature, 321:522-525 (1986); Reichmann, L., et al, Nature, 332:323-327(1988); Verhoeyen, M, et al, Science, 239:1534-36 (1988), thedisclosures of each of which are herein incorporated by reference intheir entireties).

Antibody polypeptides of the invention having PCSK9 binding specificitymay also be produced by constructing, using conventional techniques wellknown to those of ordinary skill in the art, an expression vectorcontaining an operon and a DNA sequence encoding an antibody heavy chainin which the DNA sequence encoding the CDRs required for antibodyspecificity is derived from a non-human cell source, preferably a rabbitB-cell source, while the DNA sequence encoding the remaining parts ofthe antibody chain is derived from a human cell source.

A second expression vector is produced using the same conventional meanswell known to those of ordinary skill in the art, said expression vectorcontaining an operon and a DNA sequence encoding an antibody light chainin which the DNA sequence encoding the CDRs required for antibodyspecificity is derived from a non-human cell source, preferably a rabbitB-cell source, while the DNA sequence encoding the remaining parts ofthe antibody chain is derived from a human cell source.

The expression vectors are transfected into a host cell by conventiontechniques well known to those of ordinary skill in the art to produce atransfected host cell, said transfected host cell cultured byconventional techniques well known to those of ordinary skill in the artto produce said antibody polypeptides.

The host cell may be co-transfected with the two expression vectorsdescribed above, the first expression vector containing DNA encoding anoperon and a light chain-derived polypeptide and the second vectorcontaining DNA encoding an operon and a heavy chain-derived polypeptide.The two vectors contain different selectable markers, but preferablyachieve substantially equal expression of the heavy and light chainpolypeptides. Alternatively, a single vector may be used, the vectorincluding DNA encoding both the heavy and light chain polypeptides. Thecoding sequences for the heavy and light chains may comprise cDNA,genomic DNA, or both.

Host cells which potentially may be used to express the subject antibodypolypeptides may include bacterial cells such as E. coli, or eukaryoticcells such as P. pastoris, other yeast cells, fungi, insect cells,mammalian cells, and plant cells. In one embodiment of the invention, amammalian cell of a well-defined type may be for this purpose, such as amyeloma cell, a Chinese hamster ovary (CHO) cell line, a NSO cell line,or a HEK293 cell line.

The general methods by which the vectors may be constructed,transfection methods required to produce the host cell and culturingmethods required to produce the antibody polypeptides from said hostcells all include conventional techniques. Although preferably the cellline used to produce the antibody is a mammalian cell line, any othersuitable cell line, such as a bacterial cell line such as an E.coli-derived bacterial strain, or a yeast cell line, may alternativelybe used.

Similarly, once produced the antibody polypeptides may be purifiedaccording to standard procedures in the art, such as for examplecross-flow filtration, ammonium sulphate precipitation, affinity columnchromatography and the like.

The antibody polypeptides described herein may also be used for thedesign and synthesis of either peptide or non-peptide mimetics thatwould be useful for the same therapeutic applications as the antibodypolypeptides of the invention. See, for example, Saragobi et al,Science, 253:792-795 (1991), the contents of which is hereinincorporated by reference in its entirety.

Screening Assays

The invention also includes screening assays designed to assist in theidentification of diseases and disorders associated with PCSK9 inpatients exhibiting symptoms of a PCSK9 associated disease or disorder.

In some embodiments, the antibody is used as a diagnostic tool. Theantibody can be used to assay the amount of PCSK9 present in a sampleand/or subject. As will be appreciated by one of skill in the art, suchantibodies need not be neutralizing antibodies. In some embodiments, thediagnostic antibody is not a neutralizing antibody. In some embodiments,the diagnostic antibody binds to a different epitope than theneutralizing antibody binds to. In some embodiments, the two antibodiesdo not compete with one another.

In some embodiments, the antibodies disclosed herein are used orprovided in an assay kit and/or method for the detection of PCSK9 inmammalian tissues or cells in order to screen/diagnose for a disease ordisorder associated with changes in levels of PCSK9. The kit comprisesan antibody that binds PCSK9 and means for indicating the binding of theantibody with PCSK9, if present, and optionally PCSK9 protein levels.Various means for indicating the presence of an antibody can be used.For example, fluorophores, other molecular probes, or enzymes can belinked to the antibody and the presence of the antibody can be observedin a variety of ways. The method for screening for such disorders caninvolve the use of the kit, or simply the use of one of the disclosedantibodies and the determination of whether the antibody binds to PCSK9in a sample. As will be appreciated by one of skill in the art, high orelevated levels of PCSK9 will result in larger amounts of the antibodybinding to PCSK9 in the sample. Thus, degree of antibody binding can beused to determine how much PCSK9 is in a sample. Subjects or sampleswith an amount of PCSK9 that is greater than a predetermined amount(e.g., an amount or range that a person without a PCSK9 related disorderwould have) can be characterized as having a PCSK9 mediated disorder. Insome embodiments, the antibody is administered to a subject taking astatin, in order to determine if the statin has affected the amount ofPCSK9 in the subject.

The invention is also directed to a method of in vivo imaging whichdetects the presence of cells which express PCSK9 comprisingadministering a diagnostically effective amount of a diagnosticcomposition. Said in vivo imaging is useful for the detection or imagingof PCSK9 expressing cells or organs, for example, and can be useful aspart of a planning regimen for the design of an effective cancertreatment protocol. The treatment protocol may include, for example, oneor more of radiation, chemotherapy, cytokine therapy, gene therapy, andantibody therapy, as well as an anti-PCSK9 antibody or fragment thereof.

The present invention further provides for a kit for detecting bindingof an anti-PCSK9 antibody of the invention to PCSK9. In particular, thekit may be used to detect the presence of a PCSK9 specifically reactivewith an anti-PCSK9 antibody of the invention or an immunoreactivefragment thereof. The kit may also include an antibody bound to asubstrate, a secondary antibody reactive with the antigen and a reagentfor detecting a reaction of the secondary antibody with the antigen.Such a kit may be an ELISA kit and can comprise the substrate, primaryand secondary antibodies when appropriate, and any other necessaryreagents such as detectable moieties, enzyme substrates, and colorreagents, for example as described herein. The diagnostic kit may alsobe in the form of an immunoblot kit. The diagnostic kit may also be inthe form of a chemiluminescent kit (Meso Scale Discovery, Gaithersburg,Md.). The diagnostic kit may also be a lanthanide-based detection kit(PerkinElmer, San Jose, Calif.).

A skilled clinician would understand that a biological sample includes,but is not limited to, sera, plasma, urine, saliva, mucous, pleuralfluid, synovial fluid and spinal fluid.

Methods of Ameliorating or Reducing Symptoms of or Treating, orPreventing, Diseases and Disorders Associated with, PCSK9

In another embodiment of the invention, anti-PCSK9 antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, diseases and disordersassociated with PCSK9. Anti-PCSK9 antibodies described herein, orfragments thereof, as well as combinations, can also be administered ina therapeutically effective amount to patients in need of treatment ofdiseases and disorders associated with PCSK9 in the form of apharmaceutical or diagnostic composition as described in greater detailbelow.

In another embodiment of the invention, anti-PCSK9 antibodies describedherein, or fragments thereof, with or without a second agent, are usefulfor ameliorating or reducing the symptoms of, or treating, orpreventing, disorders that relate to, involve, or can be influenced byvaried cholesterol, LDL, or LDLR levels. In some embodiments, a“cholesterol related disorder” (which includes “serum cholesterolrelated disorders”) includes any one or more of the following:hypercholesterolemia, heart disease, metabolic syndrome, diabetes,coronary heart disease, stroke, cardiovascular diseases, Alzheimer'sdisease and generally dyslipidemias, which can be manifested, forexample, by an elevated total serum cholesterol, elevated LDL, elevatedtriglycerides, elevated VLDL, and/or low HDL. Some non-limiting examplesof primary and secondary dyslipidemias that can be treated using anantibody or antibody fragment according to the invention, either alone,or in combination with one or more other agents include the metabolicsyndrome, diabetes mellitus, familial combined hyperlipidemia, familialhypertriglyceridemia, familial hypercholesterolemias, includingheterozygous hypercholesterolemia, homozygous hypercholesterolemia,familial defective apoplipoprotein B-100; polygenichypercholesterolemia; remnant removal disease, hepatic lipasedeficiency; dyslipidemia secondary to any of the following: dietaryindiscretion, hypothyroidism, drugs including estrogen and progestintherapy, beta-blockers, and thiazide diuretics; nephrotic syndrome,chronic renal failure, Cushing's syndrome, primary biliary cirrhosis,glycogen storage diseases, hepatoma, cholestasis, acromegaly,insulinoma, isolated growth hormone deficiency, and alcohol-inducedhypertriglyceridemia. antibody or antibody fragment according to theinvention can also be useful in preventing or treating atheroscleroticdiseases, such as, for example, coronary heart disease, coronary arterydisease, peripheral arterial disease, stroke (ischaemic andhemorrhagic), angina pectoris, or cerebrovascular disease and acutecoronary syndrome, myocardial infarction. In some embodiments, theantibody or antibody fragment according to the invention is useful inreducing the risk of: nonfatal heart attacks, fatal and non-fatalstrokes, certain types of heart surgery, hospitalization for heartfailure, chest pain in patients with heart disease, and/orcardiovascular events because of established heart disease such as priorheart attack, prior heart surgery, and/or chest pain with evidence ofclogged arteries. In some embodiments, the antibody or antibody fragmentaccording to the invention and methods can be used to reduce the risk ofrecurrent cardiovascular events

Administration

In one embodiment of the invention, the anti-PCSK9 antibodies describedherein, or PCSK9 binding fragments thereof, as well as combinations ofsaid antibodies or antibody fragments, are administered to a subject ata concentration of between about 0.1 and 100.0 mg/kg of body weight ofrecipient subject. In a preferred embodiment of the invention, theanti-PCSK9 antibodies described herein, or PCSK9 binding fragmentsthereof, as well as combinations of said antibodies or antibodyfragments, are administered to a subject at a concentration of about 0.4mg/kg of body weight of recipient subject. In a preferred embodiment ofthe invention, the anti-PCSK9 antibodies described herein, or PCSK9binding fragments thereof, as well as combinations of said antibodies orantibody fragments, are administered to a recipient subject with afrequency of once every twenty-six weeks or less, such as once everysixteen weeks or less, once every eight weeks or less, once every fourweeks or less, once every two weeks or less, once every week or less, oronce daily or less.

Fab fragments may be administered every two weeks or less, every week orless, once daily or less, multiple times per day, and/or every fewhours. In one embodiment of the invention, a patient receives Fabfragments of 0.1 mg/kg to 40 mg/kg per day given in divided doses of 1to 6 times a day, or in a sustained release form, effective to obtaindesired results.

It is to be understood that the concentration of the antibody or Fabadministered to a given patient may be greater or lower than theexemplary administration concentrations set forth above.

A person of skill in the art would be able to determine an effectivedosage and frequency of administration through routine experimentation,for example guided by the disclosure herein and the teachings inGoodman, L. S., Gilman, A., Brunton, L. L., Lazo, J. S., & Parker, K. L.(2006). Goodman & Gilman's the pharmacological basis of therapeutics.New York: McGraw-Hill; Howland, R. D., Mycek, M. J., Harvey, R. A.,Champe, P. C., & Mycek, M. J. (2006). Pharmacology. Lippincott'sillustrated reviews. Philadelphia: Lippincott Williams & Wilkins; andGolan, D. E. (2008). Principles of pharmacology: the pathophysiologicbasis of drug therapy. Philadelphia, Pa., [etc.]: Lippincott Williams &Wilkins.

In another embodiment of the invention, the anti-PCSK9 antibodiesdescribed herein, or PCSK9 binding fragments thereof, as well ascombinations of said antibodies or antibody fragments, are administeredto a subject in a pharmaceutical formulation.

A “pharmaceutical composition” refers to a chemical or biologicalcomposition suitable for administration to a mammal. Such compositionsmay be specifically formulated for administration via one or more of anumber of routes, including but not limited to buccal, epicutaneous,epidural, inhalation, intraarterial, intracardial,intracerebroventricular, intradermal, intramuscular, intranasal,intraocular, intraperitoneal, intraspinal, intrathecal, intravenous,oral, parenteral, rectally via an enema or suppository, subcutaneous,subdermal, sublingual, transdermal, and transmucosal. In addition,administration can occur by means of injection, powder, liquid, gel,drops, or other means of administration.

In one embodiment of the invention, the anti-PCSK9 antibodies describedherein, or PCSK9 binding fragments thereof, as well as combinations ofsaid antibodies or antibody fragments, may be optionally administered incombination with one or more active agents. Such active agents includeanalgesic, anti-histamine, antipyretic, anti-inflammatory, antibiotic,antiviral, and anti-cytokine agents. Active agents include agonists,antagonists, and modulators of TNF-α, IL-2, IL-4, IL-6, IL-10, IL-12,IL-13, IL-18, IFN-α, IFN-γ, BAFF, CXCL13, IP-10, VEGF, EPO, EGF, HRG,Hepatocyte Growth Factor (HGF), Hepcidin, including antibodies reactiveagainst any of the foregoing, and antibodies reactive against any oftheir receptors. Active agents also include but are not limited to2-Arylpropionic acids, Aceclofenac, Acemetacin, Acetylsalicylic acid(Aspirin), Alclofenac, Alminoprofen, Amoxiprin, Ampyrone, Arylalkanoicacids, Azapropazone, Benorylate/Benorilate, Benoxaprofen, Bromfenac,Carprofen, Celecoxib, Choline magnesium salicylate, Clofezone, COX-2inhibitors, Dexibuprofen, Dexketoprofen, Diclofenac, Diflunisal,Droxicam, Ethenzamide, Etodolac, Etoricoxib, Faislamine, fenamic acids,Fenbufen, Fenoprofen, Flufenamic acid, Flunoxaprofen, Flurbiprofen,Ibuprofen, Ibuproxam, Indometacin, Indoprofen, Kebuzone, Ketoprofen,Ketorolac, Lornoxicam, Loxoprofen, Lumiracoxib, Magnesium salicylate,Meclofenamic acid, Mefenamic acid, Meloxicam, Metamizole, Methylsalicylate, Mofebutazone, Nabumetone, Naproxen, N-Arylanthranilic acids,Nerve Growth Factor (NGF), Oxametacin, Oxaprozin, Oxicams,Oxyphenbutazone, Parecoxib, Phenazone, Phenylbutazone, Phenylbutazone,Piroxicam, Pirprofen, profens, Proglumetacin, Pyrazolidine derivatives,Rofecoxib, Salicyl salicylate, Salicylamide, Salicylates, Substance P,Sulfinpyrazone, Sulindac, Suprofen, Tenoxicam, Tiaprofenic acid,Tolfenamic acid, Tolmetin, and Valdecoxib.

An anti-histamine can be any compound that opposes the action ofhistamine or its release from cells (e.g., mast cells). Anti-histaminesinclude but are not limited to acrivastine, astemizole, azatadine,azelastine, betatastine, brompheniramine, buclizine, cetirizine,cetirizine analogues, chlorpheniramine, clemastine, CS 560,cyproheptadine, desloratadine, dexchlorpheniramine, diphenhydramine,ebastine, epinastine, fexofenadine, HSR 609, hydroxyzine, levocabastine,loratidine, methscopolamine, mizolastine, norastemizole, phenindamine,promethazine, pyrilamine, terfenadine, and tranilast.

Antibiotics include but are not limited to Amikacin, Aminoglycosides,Amoxicillin, Ampicillin, Ansamycins, Arsphenamine, Azithromycin,Azlocillin, Aztreonam, Bacitracin, Carbacephem, Carbapenems,Carbenicillin, Cefaclor, Cefadroxil, Cefalexin, Cefalothin, Cefalotin,Cefamandole, Cefazolin, Cefdinir, Cefditoren, Cefepime, Cefixime,Cefoperazone, Cefotaxime, Cefoxitin, Cefpodoxime, Cefprozil,Ceftazidime, Ceftibuten, Ceftizoxime, Ceftobiprole, Ceftriaxone,Cefuroxime, Cephalosporins, Chloramphenicol, Cilastatin, Ciprofloxacin,Clarithromycin, Clindamycin, Cloxacillin, Colistin, Co-trimoxazole,Dalfopristin, Demeclocycline, Dicloxacillin, Dirithromycin, Doripenem,Doxycycline, Enoxacin, Ertapenem, Erythromycin, Ethambutol,Flucloxacillin, Fosfomycin, Furazolidone, Fusidic acid, Gatifloxacin,Geldanamycin, Gentamicin, Glycopeptides, Herbimycin, Imipenem,Isoniazid, Kanamycin, Levofloxacin, Lincomycin, Linezolid, Lomefloxacin,Loracarbef, Macrolides, Mafenide, Meropenem, Meticillin, Metronidazole,Mezlocillin, Minocycline, Monobactams, Moxifloxacin, Mupirocin,Nafcillin, Neomycin, Netilmicin, Nitrofurantoin, Norfloxacin, Ofloxacin,Oxacillin, Oxytetracycline, Paromomycin, Penicillin, Penicillins,Piperacillin, Platensimycin, Polymyxin B, Polypeptides, Prontosil,Pyrazinamide, Quinolones, Quinupristin, Rifampicin, Rifampin,Roxithromycin, Spectinomycin, Streptomycin, Sulfacetamide,Sulfamethizole, Sulfanilimide, Sulfasalazine, Sulfisoxazole,Sulfonamides, Teicoplanin, Telithromycin, Tetracycline, Tetracyclines,Ticarcillin, Timidazole, Tobramycin, Trimethoprim,Trimethoprim-Sulfamethoxazole, Troleandomycin, Trovafloxacin, andVancomycin.

Active agents also include Aldosterone, Beclometasone, Betamethasone,Corticosteroids, Cortisol, Cortisone acetate, Deoxycorticosteroneacetate, Dexamethasone, Fludrocortisone acetate, Glucocorticoids,Hydrocortisone, Methylprednisolone, Prednisolone, Prednisone, Steroids,and Triamcinolone. Any suitable combination of these active agents isalso contemplated.

A “pharmaceutical excipient” or a “pharmaceutically acceptableexcipient” is a carrier, usually a liquid, in which an activetherapeutic agent is formulated. In one embodiment of the invention, theactive therapeutic agent is a humanized antibody described herein, orone or more fragments thereof. The excipient generally does not provideany pharmacological activity to the formulation, though it may providechemical and/or biological stability, and release characteristics.Exemplary formulations can be found, for example, in Remington'sPharmaceutical Sciences, 19^(th) Ed., Grennaro, A., Ed., 1995 which isincorporated by reference.

As used herein “pharmaceutically acceptable carrier” or “excipient”includes any and all solvents, dispersion media, coatings, antibacterialand antifungal agents, isotonic and absorption delaying agents that arephysiologically compatible. In one embodiment, the carrier is suitablefor parenteral administration. Alternatively, the carrier can besuitable for intravenous, intraperitoneal, intramuscular, or sublingualadministration. Pharmaceutically acceptable carriers include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. The use of such media and agents for pharmaceuticallyactive substances is well known in the art. Except insofar as anyconventional media or agent is incompatible with the active compound,use thereof in the pharmaceutical compositions of the invention iscontemplated. Supplementary active compounds can also be incorporatedinto the compositions.

Pharmaceutical compositions typically must be sterile and stable underthe conditions of manufacture and storage. The invention contemplatesthat the pharmaceutical composition is present in lyophilized form. Thecomposition can be formulated as a solution, microemulsion, liposome, orother ordered structure suitable to high drug concentration. The carriercan be a solvent or dispersion medium containing, for example, water,ethanol, polyol (for example, glycerol, propylene glycol, and liquidpolyethylene glycol), and suitable mixtures thereof. The inventionfurther contemplates the inclusion of a stabilizer in the pharmaceuticalcomposition. The proper fluidity can be maintained, for example, by themaintenance of the required particle size in the case of dispersion andby the use of surfactants.

In many cases, it will be preferable to include isotonic agents, forexample, sugars, polyalcohols such as mannitol or sorbitol, or sodiumchloride in the composition. Prolonged absorption of the injectablecompositions can be brought about by including in the composition anagent which delays absorption, for example, monostearate salts andgelatin. Moreover, the alkaline polypeptide can be formulated in a timerelease formulation, for example in a composition which includes a slowrelease polymer. The active compounds can be prepared with carriers thatwill protect the compound against rapid release, such as a controlledrelease formulation, including implants and microencapsulated deliverysystems. Biodegradable, biocompatible polymers can be used, such asethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, polylactic acid and polylactic, polyglycolic copolymers(PLG). Many methods for the preparation of such formulations are knownto those skilled in the art.

For each of the recited embodiments, the compounds can be administeredby a variety of dosage forms. Any biologically-acceptable dosage formknown to persons of ordinary skill in the art, and combinations thereof,are contemplated. Examples of such dosage forms include, withoutlimitation, reconstitutable powders, elixirs, liquids, solutions,suspensions, emulsions, powders, granules, particles, microparticles,dispersible granules, cachets, inhalants, aerosol inhalants, patches,particle inhalants, implants, depot implants, injectables (includingsubcutaneous, intramuscular, intravenous, and intradermal), infusions,and combinations thereof.

The above description of various illustrated embodiments of theinvention is not intended to be exhaustive or to limit the invention tothe precise form disclosed. While specific embodiments of, and examplesfor, the invention are described herein for illustrative purposes,various equivalent modifications are possible within the scope of theinvention, as those skilled in the relevant art will recognize. Theteachings provided herein of the invention can be applied to otherpurposes, other than the examples described above.

These and other changes can be made to the invention in light of theabove detailed description. In general, in the following claims, theterms used should not be construed to limit the invention to thespecific embodiments disclosed in the specification and the claims.Accordingly, the invention is not limited by the disclosure, but insteadthe scope of the invention is to be determined entirely by the followingclaims.

The invention may be practiced in ways other than those particularlydescribed in the foregoing description and examples. Numerousmodifications and variations of the invention are possible in light ofthe above teachings and, therefore, are within the scope of the appendedclaims.

Certain teachings related to humanization of rabbit-derived monoclonalantibodies and preferred sequence modifications to maintain antigenbinding affinity were disclosed in International Application No.PCT/US2008/064421, corresponding to International Publication No.WO/2008/144757, entitled “Novel Rabbit Antibody Humanization Methods andHumanized Rabbit Antibodies”, filed May 21, 2008, the disclosure ofwhich is herein incorporated by reference in its entirety.

Certain teachings related to producing antibodies or fragments thereofusing mating competent yeast and corresponding methods were disclosed inU.S. patent application Ser. No. 11/429,053, filed May 8, 2006, (U.S.Patent Application Publication No. US2006/0270045), the disclosure ofwhich is herein incorporated by reference in its entirety.

Certain PCSK9 antibody polynucleotides and polypeptides are disclosed inthe sequence listing accompanying this patent application filing, andthe disclosure of said sequence listing is herein incorporated byreference in its entirety.

The entire disclosure of each document cited (including patents, patentapplications, journal articles, abstracts, manuals, books, or otherdisclosures) in the Background of the Invention, Detailed Description,and Examples is herein incorporated by reference in their entireties.

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the subject invention, and are not intended to limit thescope of what is regarded as the invention. Efforts have been made toensure accuracy with respect to the numbers used (e.g. amounts,temperature, concentrations, etc.) but some experimental errors anddeviations should be allowed for. Unless otherwise indicated, parts areparts by weight, molecular weight is average molecular weight,temperature is in degrees centigrade; and pressure is at or nearatmospheric.

EXAMPLES Example 1 Preparation of Antibodies that Bind PCSK9

By using an antibody selection protocol substantially as describedherein, a panel of antibodies specific to PCSK9 were produced.

Immunization Strategy

Rabbits were immunized with human PCSK9 (R&D and in house produced).Immunization consisted of a first subcutaneous (sc) injection of 100 μgof antigen in complete Freund's adjuvant (CFA) (Sigma) followed by twoboosts, two weeks apart each containing 50 μg antigen mixed with 50 μgin incomplete Freund's adjuvant (IFA) (Sigma). Animals were bled on day55, and serum titers were determined by ELISA (antigen recognition).

Additional immunizations were performed with repetitive immunizations inmultiple sites. Animals were given either a high or low dose of antigenon the following schedule: initial dose (400 μg or 100 μg) on Day 1 withboosts (200 μg or 50 μg) on Days 3, 6, 9, and 12. Tissue harvest wasperformed on Day 13.

Antibody Selection Titer Assessment

To identify and characterize antibodies that bind to human PCSK9,antibody-containing solutions were tested by ELISA. Briefly, neutravidincoated plates (Thermo Scientific), were coated with biotinylated humanPCSK9 (504, per well, 1 μg/mL) diluted in PBS for approximately 1 hr atroom temperature or alternatively overnight at 4° C. The plates werethen blocked with ELISA buffer for one hour at room temperature andwashed using wash buffer (PBS, 0.05% tween 20). Serum samples testedwere serially diluted using ELISA buffer (0.5% fish skin gelatin in PBSpH 7.4). Fifty microliters of diluted serum samples were transferredonto the wells and incubated for one hour at room temperature for onehour. After this incubation, the plate was washed with wash buffer. Fordevelopment, an anti-rabbit specific Fc-HRP (1:5000 dilution in ELISAbuffer) was added onto the wells and incubated for 45 min at RT. After a3× wash step with wash solution, the plate was developed using TMBsubstrate for two minutes at room temperature and the reaction wasquenched using 0.5M HCl. The well absorbance was read at 450 nm.

Tissue Harvesting

Once acceptable titers were established, the rabbit(s) were sacrificed.Spleen, lymph nodes, and whole blood were harvested and processed asfollows:

Spleen and lymph nodes were processed into a single cell suspension bydisassociating the tissue and pushing through sterile wire mesh at 70 μm(Fisher) with a plunger of a 20 cc syringe. Cells were collected in PBS.Cells were washed twice by centrifugation. After the last wash, celldensity was determined by trypan blue. Cells were centrifuged at 1500RPM for 10 minutes; the supernatant was discarded. Cells wereresuspended in the appropriate volume of 10% dimethyl sulfoxide (DMSO,Sigma) in FBS (Hyclone) and dispensed at 1 mL/vial. Vials were stored at−70° C. in a slow freezing chamber for 24 hours and stored in liquidnitrogen.

B Cell Selection, Enrichment and Culture Conditions

On the day of setting up B cell cultures, PBMC, splenocyte, or lymphnode vials were thawed for use. Vials were removed from LN2 tank andplaced in a 37° C. water bath until thawed. Contents of vials weretransferred into 15 mL conical centrifuge tube (Corning) and 10 mL ofmodified RPMI described above was slowly added to the tube. Cells werecentrifuged for 5 minutes at 2K RPM, and the supernatant was discarded.Cells were resuspended in 10 mL of fresh media. Cell density andviability was determined by trypan blue.

Biotinylated human PCSK9 was pre-loaded onto the streptavidin beads asfollows. Seventy five microliters of streptavidin beads (Milteny Biotec,Auburn Calif.) were mixed with N-terminally biotinylated huPCSK9 (10μg/mL final concentration) and 300 μl PBF. This mixture was incubated at4° C. for 30 min and unbound biotinylated human PCSK9 was removed usinga MACS® separation column (Miltenyi Biotec, with a 1 ml rinse to removeunbound material. Then material was plunged out, then used to resuspendcells from above in 100 μL per 1E7 cells, the mixture was then incubatedat 4° C. for 30 min and washed once with 10 mL of PBF. After washing,the cells were resuspended in 500 μL of PBF and set aside. A MACS® MScolumn (Miltenyi Biotec, Auburn Calif.) was pre-rinsed with 500 mL ofPBF on a magnetic stand (Milteni). Cell suspension was applied to thecolumn through a pre-filter, and unbound fraction was collected. Thecolumn was washed with 2.5 mL of PBF buffer. The column was removed fromthe magnet stand and placed onto a clean, sterile 1.5 mL eppendorf tube.1 mL of PBF buffer was added to the top of the column, and positiveselected cells were collected. The yield and viability of positive cellfraction was determined by trypan blue staining. Positive selectionyielded an average of 1% of the starting cell concentration.

A pilot cell screen was established to provide information on seedinglevels for the culture. Plates were seeded at 10, 25, 50, 100, or 200enriched B cells/well. In addition, each well contained 25-50Kcells/well of irradiated EL-4.B5 cells (5,000 Rads) and an appropriatelevel of activated rabbit T cell supernatant (See U.S. PatentApplication Publication No. 20070269868) (ranging from 1-5% depending onpreparation) in high glucose modified RPMI medium at a final volume of250 μL/well. Cultures were incubated for 5 to 7 days at 37° C. in 4%CO₂.

B-Cell Culture Screening by Antigen-Recognition (ELISA)

To identify wells producing anti-human PCSK9 antibodies, the sameprotocol as described for titer determination of serum samples byantigen-recognition (ELISA) was used with the following changes.Briefly, neutravidin coated plates were coated with biotinylated humanPCSK9 (504 per well, 1 μg/mL each). B-cell supernatant samples (50 μL)were tested without prior dilution.

Identification of Functional Activity in B-Cell Supernatants Using Oneor More Assays

Following identification by antigen-recognition ELISA, antibodiestargeting the function modifying epitopes of PCSK9 were identified usinga PCSK9 blocking ELISA. Biotinylated PCSK9 was coated on to streptavidinplates and then a pool of recombinant, function modifying anti-PCSK9antibodies (in house generated) with human Fc regions were used to blockfunctional epitopes. Once the epitopes were blocked, B-cell culturesupernatants were put on the wells and antibody binding detected byanti-rabbit Fc-HRP (Jackson ImmunoResearch).

Amplification and Sequence Determination of Antibody Sequences fromAntigen-Specific B Cells

After effecting functional assays to identify B cell supernatantscontaining anti-PCSK9 antibodies possessing desired functionalproperties, antigen-specific B cells secreting these antibodies werethen isolated. The antibody sequences were thereafter recovered fromthese isolated B cells using a combined RT-PCR based method thatisolates antibody sequences from a single isolated B-cell. Primerscontaining restriction enzyme recognition sites were designed to annealin conserved and constant regions of the target immunoglobulin genes(heavy and light), such as rabbit immunoglobulin sequences, and atwo-step nested PCR recovery was used to amplify the antibody sequence.Amplicons from each well were analyzed for recovery and size integrity.The resulting fragments are then digested with AluI to fingerprint thesequence clonality. Identical sequences displayed a common fragmentationpattern in their electrophoretic analysis. The original heavy and lightchain amplicon fragments were then digested using the restriction enzymesites contained within the PCR primers and cloned into an expressionvector. Vector containing subcloned DNA fragments were amplified andpurified. Sequence of the subcloned heavy and light chains were verifiedprior to expression.

Recombinant Production of Monoclonal Antibody of Desired AntigenSpecificity and/or Functional Properties

To determine antigen specificity and functional properties of recoveredantibodies from specific B-cells, vectors driving the expression of thedesired paired heavy and light chain sequences were transfected intoHEK-293 cells.

Antigen-Recognition of Recombinant Antibodies by ELISA

To characterize recombinant expressed antibodies for their ability tobind to human PCSK9, antibody-containing solutions were tested by ELISA.All incubations were done at room temperature. Briefly, Neutravidinplates (Pierce) were blocked for 1 hr with ELISA buffer (PBS, 0.5% fishskin gelatin, 0.05% Tween-20). After blocking, plates were coated with abiotinylated-PCSK9 containing solution (1 μg/mL in ELISA buffer) for 1hour. PCSK9-coated plates were then washed three times in wash buffer(PBS, 0.05% Tween-20). After coating, the plates were blocked again withELISA buffer for 1 hour. The blocking solution was removed and theplates were then incubated with a dilution series of the antibody beingtested for approximately one hour. At the end of this incubation, theplate was washed three times with wash buffer and further incubated witha secondary antibody containing solution (Peroxidase conjugatedaffinipure F(ab′)2 fragment goat anti-human IgG, Fc fragment specific[Jackson Immunoresearch]) for approximately 45 minutes and washed threetimes. Next a substrate solution (TMB peroxidase substrate, BioFx) wasadded and incubated for 3 to 5 minutes in the dark. The reaction wasstopped by addition of 0.5M HCl and the plate was read at 450 nm in aplate-reader.

Results: FIGS. 13-26 demonstrate that anti-PCSK9 antibodies Ab1-Ab24bind to and recognize human PCSK9.

Functional Characterization of Recombinant Antibodies by Modulation ofLDL-C Uptake by HepG2 Cells

The ability of anti-PCSK9 antibodies to neutralize the inhibition ofLDL-C uptake in HepG2 cells by PCSK9 was tested in a cell-based assay.HepG2 cells were seeded (30,000 cells/well) in a collagen coated 96 wellplate. Twenty-four hours later, the media was replaced with fresh mediacontaining 0.5% low lipid FBS. Various concentrations of anti-PCSK9antibodies were incubated with 3 μg/mL PCSK9 for 1 hour at roomtemperature and then added to the HepG2 cells and incubated for 5 hoursat 37° C. BODIPY-LDL was added to each well and incubated overnight at37° C. The media was removed and the cells lysed with RIPA buffer andthe amount of BODIPY-LDL taken up by the cells measured on a platereader (excitation, 485 nm; emission 535 nm).

Results: FIGS. 27-46 demonstrate that anti-PCSK9 antibodies neutralizethe ability of PCSK9 to inhibit LDL-C uptake.

Cynomolgus Monkey Antigen-Recognition of Recombinant Antibodies by ELISA

To characterize recombinant expressed antibodies for their ability tobind to cynomolgus monkey PCSK9, antibody-containing solutions weretested by ELISA. Procedure was as described for human PCSK9 ELISA exceptcynomolgus PCSK9 was utilized.

Results: FIGS. 47-60 demonstrate that anti-PCSK9 antibodies bind andrecognize cynomolgus monkey PCSK9.

Example 2 Yeast Cell Expression

Construction of Pichia pastoris Expression Vectors for Heavy and LightChain

The humanized light and heavy chain fragments were commerciallysynthesized and subcloned into a pGAP expression vector. The pGAPexpression vector uses the GAP promoter to drive expression of theimmunoglobulin chain and a secretion leader sequence for export. Inaddition, this vector contains common elements such as a bacterialorigin of replication, and a copy of the kanamycin resistance gene whichconfers resistance to the antibiotic G418 in P. pastoris. G418 providesa means of selection for strains that contain the desired expressionvector integrated into their genome.

Transformation of Expression Vectors into Haploid Met1 and Lys3 HostStrains of Pichia pastoris

All methods used for transformation of haploid P. pastoris strains andmanipulation of the P. pastoris sexual cycle were done as described inPichia Protocols (Methods in Molecular Biology Higgings, D R, and Cregg,J M, Eds. 1998. Humana Press, Totowa, N.J.). Prior to transformationeach vector was linearized within the GAP promoter sequences to directthe integration of the vector into the GAP promoter locus of the P.pastoris genome. Haploid strains were transfected using electroporationand successful transformants were selected on YPDS (yeast extract,peptone dextrose with sorbitol) G418 agar plates. Copy numbers of heavyand light chain genes were determined for haploid strains by Southernblot analysis. Haploid strains were then mated and selected for theirability to grow in the absence of the amino acid markers (i.e., Lys andMet). Resulting diploid clones were then subjected to a final Southernblot to confirm copy numbers of heavy and light chain genes. A cloneexpressing the antibody of interest was selected using biolayerinterferometry Protein-A biosensors to monitor expression (Octet,ForteBio). Dual locus strains were generated using the methods disclosedin U.S. Application No. 61/525,307, the contents of which areincorporated by reference in its entirety.

Example 3 Expression of Ab18 in Pichia pastoris

Pichia strains for expression of full-length antibody were made. For allthe full length antibody expressing strains, haploids strains werecreated and subsequently mated. One haploid strain expressed full-lengthlight chain sequence and another haploid strain expressed thefull-length heavy chain sequence. Each diploid strain was used togenerate a research cell bank and used for expression in a bioreactor.

First an inoculum was expanded using the research cell bank using mediumcomprised of the following nutrients (% w/v): yeast extract 3%,anhydrous dextrose 4%, YNB 1.34%, Biotin 0.004% and 100 mM potassiumphosphate. To generate the inoculum for the fermenters, the cell bankwas expanded for approximately 24 hours in a shaking incubator at 30° C.and 300 RPM. A 10% inoculum was then added to Labfors 2.5 L workingvolume vessels containing 1 L sterile growth medium. The growth mediumwas comprised of the following nutrients: potassium sulfate 18.2 g/L,ammonium phosphate monobasic 36.4 g/L, potassium phosphate dibasic 12.8g/L, magnesium sulfate heptahydrate 3.72 g/L, sodium citrate dihydrate10 g/L, glycerol 40 g/L, yeast extract 30 g/L, PTM1 trace metals 4.35mL/L, and antifoam 204 1.67 mL/L. The PTM1 trace metal solution wascomprised of the following components: cupric sulfate pentahydrate 6g/L, sodium iodide 0.08 g/L, manganese sulfate hydrate 3 g/L, sodiummolybdate dehydrate 0.2 g/L, boric acid 0.02 g/L, cobalt chloride 0.5g/L, zinc chloride 20 g/L, ferrous sulfate heptahydrate 65 g/L, biotin0.2 g/L, and sulfuric acid 5 mL/L.

The bioreactor process control parameters were set as follows: Agitation1,000 RPM, airflow 1.35 standard liter per minute, temperature 28° C.and pH was controlled at six using ammonium hydroxide. No oxygensupplementation was provided.

Fermentation cultures were grown for approximately 12 to 16 hours untilthe initial glycerol was consumed as denoted by a dissolved oxygenspike. The cultures were starved for approximately three hours after thedissolved oxygen spike. After this starvation period, a bolus additionof ethanol was added to the reactor to reach 1% ethanol (w/v). Thefermentation cultures were allowed to equilibrate for 15 to 30 minutes.Feed addition was initiated 30 minutes post-ethanol bolus and set at aconstant rate of 1 mL/min for 40 minutes, then the feed pump wascontrolled by an ethanol sensor keeping the concentration of ethanol at1% for the remainder of the run using an ethanol sensing probe (RavenBiotech). The feed was comprised of the following components: yeastextract 50 g/L, dextrose 500 g/L, magnesium sulfate heptahydrate 3 g/L,and PTM1 trace metals 12 mL/L. The total fermentation time wasapproximately 90 hours.

Example 4 Methods of Humanizing Antibodies

Methods of humanizing antibodies have been described previously inissued U.S. Pat. No. 7,935,340, the disclosure of which is incorporatedherein by reference in its entirety. In some instances, a determinationof whether additional rabbit framework residues are required to maintainactivity is necessary. In some instances the humanized antibodies stillrequires some critical rabbit framework residues to be retained tominimize loss of affinity or activity. In these cases, it is necessaryto change single or multiple framework amino acids from human germlinesequences back to the original rabbit amino acids in order to havedesired activity. These changes are determined experimentally toidentify which rabbit residues are necessary to preserve affinity andactivity.

Example 5 Binding Affinities of Anti-PCSK9 Antibodies

Binding affinities of monoclonal antibodies for PCSK9 were estimatedusing Bio-Layer Interferometry (BLI) on the Octet QK (ForteBio).Biotinylated antibody was immobilized at different densities to thesurface of Streptavidin (SA) Biosensors. A dilution series of humanPCSK9 prepared in 1× Kinetics Buffer (NaCl 0.0138 M; KCl 0.00027 M; 0.1mg/mL BSA, 0.002% Tween and 0.005% Sodium Azide; pH 7.4, at 25° C.)purchased from the manufacturer (ForteBio) was used to query theantibodies. At our chosen concentrations of antigen (˜0.25 μg/mL-2μg/mL) association times of 15 minutes and dissociation times of 25minutes were used with the Octet Analysis Software (v6.4, ForteBio), toglobally fit data using a 1:1 Langmuir binding model. This techniquedoes not allow for sensor regeneration and each antibody tested requiresa unique set of sensors. Examples of antibody affinities are listed inTable 1.

TABLE 1 Antibody K_(D) (pM) k_(on) (1/Ms) k_(dis) (1/s) Ab1  79901.62E+05 1.30E−03 Ab2  4250 1.07E+05 4.53E−04 Ab3  5870 9.44E+045.54E−04 Ab4  6000 7.03E+04 4.22E−04 Ab5  3160 8.81E+04 2.78E−04 Ab6 531 7.53E+04 4.00E−05 Ab7  379 7.07E+04 2.68E−05 Ab8  637 7.70E+044.91E−05 Ab9  637 7.70E+04 4.91E−05 Ab10 676 7.53E+04 5.09E−05 Ab11 3326.68E+04 2.22E−05 Ab12 120 1.00E+05 1.21E−05 Ab13 2230 1.59E+05 3.53E−04Ab14 3700 9.79E+04 3.62E−04 Ab15 839 6.92E+04 5.80E−05 Ab16 12807.75E+04 9.92E−05 Ab17 609 1.03E+05 6.27E−05 Ab18 215 9.70E+04 2.09E−05Ab19 1300 8.80E+04 1.16E−04 Ab20 600 1.08E+05 6.63E−05Binding Affinities of Anti-PCSK9 Antibodies Using SPR (Ab21-24)

Binding affinities of monoclonal antibodies for PCSK9 were estimatedusing Surface Plasmon Resonance (SPR) on the ProteOn XPR36 (Bio-Rad).Biotinylated antibody was immobilized at progressively increasingdensities to the surface of Neutravidin (NLC) Chips. A dilution seriesof human PCSK9 prepared in 1×HBS-EP+Buffer (10 mM Hepes; 150 mM NaCl; 3mM EDTA, 0.05% Polysorbate 20; pH 7.6 at 25° C.) purchased from ThermoScientific and supplemented with 0.2 mg/mL Bovine Serum Albumin (BSA)from Jackson ImmunoResearch and 0.005% sodium azide from VWR was used toquery the antibodies. At our chosen concentrations of antigen (˜0.08μg/mL-6.7 μg/mL) association times of 4 minutes and dissociation timesof 30-120 minutes were used with the ProteOn Manager Software (v3.1.0.6,Bio-Rad) to group and fit data using a 1:1 Langmuir binding model.Surfaces were regenerated between analyte queries and after bindingantibody to the surface using 10 mM Glycine pH 2.0. Data across 4different densities were averaged and a single K_(D) and standardpropagation of error calculated for each antibody. These values arereported in Table 2.

TABLE 2 Antibody K_(D) (M) AVG hu k_(d) (1/s) AVG hu k_(a) (1/Ms) Ab216.1E−10 3.54E−04 5.80E+05 Ab22 4.9E−10 2.59E−04 5.29E+05 Ab23 4.0E−113.71E−05 9.28E+05 Ab24 9.0E−11 6.16E−05 6.84E+05

Example 6 PCSK9 Antibodies Inhibit PCSK9 Interaction with LDLR

The ability of the panel of anti-PCSK9 antibodies to block theinteraction of PCSK9 with LDLR was evaluated with an ELISA basedimmunoassay. Briefly, LDLR (0.5 m/mL) was coated on 4HBX platesovernight at 4° C. The plates were blocked with 0.5% fish skin gelatinin PBS, pH 7.4 for 1 hour at room temperature followed by 3 washes withPBS. Biotinylated PCSK9 (0.3 μg/ml) was incubated in the presence ofserial dilutions of anti-PCSK9 antibodies at room temperature for 1 hourand then added to the LDLR coated plates. After incubation for 1 hour atroom temperature, plates were washed (3× with PBS). For development,streptavidin-HRP was added, incubated for 1 hour at room temperature,washed (3× in PBS) and TMB substrated added for 3 minutes at roomtemperature. The reaction was quenched using 0.5M HCl and absorbanceread at 450 nm.

Results: FIGS. 61-84 demonstrates anti-PCSK9 antibodies block PCSK9interaction with LDLR.

Example 7 Anti-PCSK9 Antibody Lowers Plasma LDL-C Levels in Non-HumanPrimates

A pharmacodynamic (PD) and pharmacokinetic (PK) study was conducted innaïve male cynomolgus monkeys aged 3-4 years and weights of 3-4 kg.

Three cynomolgus monkeys were injected with vehicle (15 mM histidine,250 mM sorbitol, pH 5.5) and three monkeys were injected with Ab18 (5mg/kg). Injections were performed by IV bolus administered on day 1.

Blood was collected by venipuncture and processed to serum for clinicalchemistry. Blood was collected in K₃EDTA tubes and processed to plasmafor PK analyses. Blood samples were collected for clinical chemistryfrom fasted animals pre-injection, 24 hours post injection and thenevery third day to day 30. Blood samples for PK analysis were collectedpre-injection, 5 min, 30 min, 1 hr, 2 hr, 4 hr, 8 hr, 12, hr, 18 hr, 24,hr 36 hr, 48, hr, 96 hr and then every 3 days to day 30. All sampleswere stored at −70° C. or below.

Clinical chemistry analyses included the following parameters: alanineaminotransferase, aspartate aminotransferase, alkaline phosphatase,gamma-glutamyltransferase, lactate dehydrogenase, total bilirubin, ureanitrogen, C-reactive protein, calcium, creatinine, phosphorus, lowdensity lipoprotein, high density lipoprotein, total protein, albumin,globulin, albumin/globulin ratio, glucose, cholesterol, triglycerides,sodium, potassium, chloride, and bicarbonate.

Results: FIG. 85 demonstrates that Ab18 resulted in a reduction of LDL-Clevels up to 60% over a 14 day period. LDL-C levels subsequently returnrapidly to predose levels. Following injection of Ab18 on day 30 asimilar decrease on LDL-C levels was observed. A 20-25% reduction incholesterol was observed (FIG. 86) while HDL-C levels remained unchanged(FIG. 87).

Total antibody levels were determined by ELISA. Briefly, Multi-Array 96well High Bind plates were coated with 0.5 m/mL goat anti-Human IgG(monkey adsorbed) overnight at 4° C. Plates were blocked with 3% BlockerA in PBS+0.5% Tween 20 for 1 hour at room temperature. Plates werewashed (3× in PBS+0.5% Tween) and sample added followed by a 1 hourincubation at room temperature. Plates were washed 3 times and Ab18detected with biotinylated human heavy/light chain antibody andsulfo-TAG strepavadin. Plates were washed 3 times and read with MSD.

Results: Table 3 shows results in three cynomolgus monkeys of totalantibody levels

TABLE 3 Animal C_(max) (mg/ml) T_(max) (h) T_(1/2) (h) 1 72.93 4 71.35 2110.35 4 63.46 3 98.99 4 69.01

Example 8 Anti-PCSK9 Antibody Lowers Plasma LDL-C Levels in Non-HumanPrimates

A pharmacodynamic (PD) study was conducted in male cynomolgus monkeysaged 3-4 years and weights of 3-4.5 kg.

Three cynomolgus monkeys were injected with Ab11 (5 mg/kg). Injectionswere performed by IV bolus administered on day 1.

Blood was collected by venipuncture and processed to serum for clinicalchemistry. Blood was collected in K3EDTA tubes and processed to plasmafor PK analyses. Blood samples were collected for clinical chemistryfrom fasted animals pre-injection, 24 hours post injection and thenevery third day. Blood samples for PK analysis were collectedpre-injection, 5 min, 30 min, 1 hr, 2 hr, 4 hr, 8 hr, 12, hr, 18 hr, 24,hr 36 hr, 48, hr, 96 hr, 168 hr, 240 hr, 312 hr, 384 hr, 528 hr, and 696hr. All samples were stored at −70° C. or below.

Clinical chemistry analyses included the following parameters: alanineaminotransferase, aspartate aminotransferase, alkaline phosphatase,gamma-glutamyltransferase, lactate dehydrogenase, total bilirubin, ureanitrogen, C-reactive protein, calcium, creatinine, phosphorus, lowdensity lipoprotein, high density lipoprotein, total protein, albumin,globulin, albumin/globulin ratio, glucose, cholesterol, triglycerides,sodium, potassium, chloride, and bicarbonate.

Results: FIG. 88 demonstrates that Ab11 resulted in a reduction of LDL-Clevels up to 60% over a 16 day period. LDL-C levels subsequentlyreturned to predose levels. A 25-30% reduction in cholesterol wasobserved (FIG. 89) while HDL-C levels remained unchanged (FIG. 90).

Example 9 Anti-PCSK9 Antibody Lowers Plasma LDL-C Levels in Non-HumanPrimates

A pharmacodynamic (PD) study was conducted in male cynomolgus monkeysaged 3-5 years and weights of 2.8-4.0 kg.

Three cynomolgus monkeys were injected with Ab19 (5 mg/kg). Injectionswere performed by IV bolus administered on day 1.

Blood was collected by venipuncture and processed to serum for clinicalchemistry. Blood was collected in K3EDTA tubes and processed to plasmafor PK analyses. Blood samples were collected for clinical chemistryfrom fasted animals pre-injection, 24 hours, 48 hours, and 96 hours postinjection and then every third day to day 34. Blood samples for PKanalysis were collected pre-injection, 5 min, 30 min, 1 hr, 2 hr, 4 hr,8 hr, 12 hr, 24 hr, 48 hr, 96 hr, and then every third day to day 34.All samples were stored at −70° C. or below.

Clinical chemistry analyses included the following parameters: alanineaminotransferase, aspartate aminotransferase, alkaline phosphatase,gamma-glutamyltransferase, lactate dehydrogenase, total bilirubin, ureanitrogen, C-reactive protein, calcium, creatinine, phosphorus, lowdensity lipoprotein, high density lipoprotein, total protein, albumin,globulin, albumin/globulin ratio, glucose, cholesterol, triglycerides,sodium, potassium, chloride, and bicarbonate.

Results: FIG. 91 demonstrates that Ab19 resulted in a reduction of LDL-Clevels up to 65% over a 19 day period. LDL-C levels subsequentlyreturned to predose levels.

Example 10 Anti-PCSK9 Antibody Lowers Plasma LDL-C Levels in Non-HumanPrimates

A pharmacodynamic (PD) study was conducted in male cynomolgus monkeysaged 3-5 years and weights of 2.8-4.0 kg.

Three cynomolgus monkeys were injected with Ab20 (5 mg/kg). Injectionswere performed by IV bolus administered on day 1.

Blood was collected by venipuncture and processed to serum for clinicalchemistry. Blood was collected in K3EDTA tubes and processed to plasmafor PK analyses. Blood samples were collected for clinical chemistryfrom fasted animals pre-injection, 24 hours, 48 hours, and 96 hours postinjection and then every third day to day 34. Blood samples for PKanalysis were collected pre-injection, 5 min, 30 min, 1 hr, 2 hr, 4 hr,8 hr, 12 hr, 24 hr, 48 hr, 96 hr, and then every third day to day 34.All samples were stored at −70° C. or below.

Clinical chemistry analyses included the following parameters: alanineaminotransferase, aspartate aminotransferase, alkaline phosphatase,gamma-glutamyltransferase, lactate dehydrogenase, total bilirubin, ureanitrogen, C-reactive protein, calcium, creatinine, phosphorus, lowdensity lipoprotein, high density lipoprotein, total protein, albumin,globulin, albumin/globulin ratio, glucose, cholesterol, triglycerides,sodium, potassium, chloride, and bicarbonate.

Results: FIG. 92 demonstrates that Ab20 resulted in a reduction of LDL-Clevels up to 70% over a 22 day period. LDL-C levels subsequentlyreturned to predose levels.

Example 11 Anti-PCSK9 Antibody Lowers Plasma LDL-C Levels in Non-HumanPrimates

A pharmacodynamic (PD) study was conducted in male cynomolgus monkeysaged 3-5 years and weights of 2.8-4.0 kg.

Three cynomolgus monkeys were injected with Ab22 and Ab24 (5 mg/kg).Injections were performed by IV bolus administered on day 1.

Blood was collected by venipuncture and processed to serum for clinicalchemistry. Blood was collected in K3EDTA tubes and processed to plasmafor PK analyses. Blood samples were collected for clinical chemistryfrom fasted animals pre-injection, 24 hours, 48 hours, and 96 hours postinjection and then every third day to day 46 (Ab22) or day 37 (Ab24).Blood samples for PK analysis were collected pre-injection, 5 min, 30min, 1 hr, 2 hr, 4 hr, 8 hr, 12 hr, 24 hr, 48 hr, 96 hr, and then everythird day to day 46 (Ab22) or day 37 (Ab24). All samples were stored at−70° C. or below.

Clinical chemistry analyses included the following parameters: alanineaminotransferase, aspartate aminotransferase, alkaline phosphatase,gamma-glutamyltransferase, lactate dehydrogenase, total bilirubin, ureanitrogen, C-reactive protein, calcium, creatinine, phosphorus, lowdensity lipoprotein, high density lipoprotein, total protein, albumin,globulin, albumin/globulin ratio, glucose, cholesterol, triglycerides,sodium, potassium, chloride, and bicarbonate.

Results: FIG. 93 demonstrates that Ab22 resulted in a reduction of LDL-Clevels up to 60% over a 22 day period. LDL-C levels subsequentlyreturned to predose levels. FIG. 94 demonstrates that Ab24 resulted in areduction of LDL-C levels up to 70% over a 22 day period. LDL-C levelssubsequently returned to predose levels.

What is claimed is:
 1. An isolated antibody or antigen-binding fragmentthereof that binds to proprotein convertase subtilisin/kexin type 9(PCSK9), comprising a heavy chain and a light chain that each comprisecomplementarity determining regions (CDRs), wherein: (a) the variableheavy chain comprises the CDR1 sequence of SEQ ID NO: 4, the CDR2sequence of SEQ ID NO: 6, and the CDR3 sequence of SEQ ID NO: 8; and thevariable light chain comprises the CDR1 sequence of SEQ ID NO: 24, theCDR2 sequence of SEQ ID NO: 26, and the CDR3 sequence of SEQ ID NO: 28;or (b) the variable heavy chain comprises the CDR1 sequence of SEQ IDNO: 684, the CDR2 sequence of SEQ ID NO: 686, and the CDR3 sequence ofSEQ ID NO: 688; and the variable light chain comprises the CDR1 sequenceof SEQ ID NO: 704, the CDR2 sequence of SEQ ID NO: 706, and the CDR3sequence of SEQ ID NO: 708; or (c) the variable heavy chain comprisesthe CDR1 sequence of SEQ ID NO: 724, the CDR2 sequence of SEQ ID NO:726, and the CDR3 sequence of SEQ ID NO: 728; and the variable lightchain comprises the CDR1 sequence of SEQ ID NO: 744, the CDR2 sequenceof SEQ ID NO: 746, and the CDR3 sequence of SEQ ID NO: 748; or (d) thevariable heavy chain comprises the CDR1 sequence of SEQ ID NO: 764, theCDR2 sequence of SEQ ID NO: 766, and the CDR3 sequence of SEQ ID NO:768; and the variable light chain comprises the CDR1 sequence of SEQ IDNO: 784, the CDR2 sequence of SEQ ID NO: 786, and the CDR3 sequence ofSEQ ID NO: 788; or (e) the variable heavy chain comprises the CDR1sequence of SEQ ID NO: 844, the CDR2 sequence of SEQ ID NO: 846, and theCDR3 sequence of SEQ ID NO: 848; and the variable light chain comprisesthe CDR1 sequence of SEQ ID NO: 864, the CDR2 sequence of SEQ ID NO:866, and the CDR3 sequence of SEQ ID NO:
 868. 2. The antibody orantigen-binding fragment thereof of claim 1, wherein: (i) the variableheavy chain comprises a sequence at least 90% identical to SEQ ID NO: 2,the variable light chain comprises a sequence at least 90% identical toSEQ ID NO: 22, and the variable heavy and light chains comprise the CDRsaccording to (a); or (ii) the variable heavy chain comprises a sequenceat least 90% identical to SEQ ID NO: 682, the variable light chaincomprises a sequence at least 90% identical to SEQ ID NO: 702, and thevariable heavy and light chains comprise the CDRs according to (b); or(iii) the variable heavy chain comprises a sequence at least 90%identical to SEQ ID NO: 722, the variable light chain comprises asequence at least 90% identical to SEQ ID NO: 742, and the variableheavy and light chains comprise the CDRs according to (c); or (iv) thevariable heavy chain comprises a sequence at least 90% identical to SEQID NO: 762, the variable light chain comprises a sequence at least 90%identical to SEQ ID NO: 782, and the variable heavy and light chainscomprise the CDRs according to (d); or (v) the variable heavy chaincomprises a sequence at least 90% identical to SEQ ID NO: 842, thevariable light chain comprises a sequence at least 90% identical to SEQID NO: 862, and the variable heavy and light chains comprise the CDRsaccording to (e).
 3. The antibody or antigen-binding fragment thereof ofclaim 1, wherein: (i) the variable heavy chain comprises a sequence atleast 95% identical to SEQ ID NO: 2, the variable light chain comprisesa sequence at least 95% identical to SEQ ID NO: 22, and the variableheavy and light chains comprise the CDRs according to (a); or (ii) thevariable heavy chain comprises a sequence at least 95% identical to SEQID NO: 682, the variable light chain comprises a sequence at least 95%identical to SEQ ID NO: 702, and the variable heavy and light chainscomprise the CDRs according to (b); or (iii) the variable heavy chaincomprises a sequence at least 95% identical to SEQ ID NO: 722, thevariable light chain comprises a sequence at least 95% identical to SEQID NO: 742, and the variable heavy and light chains comprise the CDRsaccording to (c); or (iv) the variable heavy chain comprises a sequenceat least 95% identical to SEQ ID NO: 762, the variable light chaincomprises a sequence at least 95% identical to SEQ ID NO: 782, and thevariable heavy and light chains comprise the CDRs according to (d); or(v) the variable heavy chain comprises a sequence at least 95% identicalto SEQ ID NO: 842, the variable light chain comprises a sequence atleast 95% identical to SEQ ID NO: 862, and the variable heavy and lightchains comprise the CDRs according to (e).
 4. The antibody orantigen-binding fragment thereof of claim 1, wherein: (i) the variableheavy chain comprises SEQ ID NO: 2, the variable light chain comprisesSEQ ID NO: 22, and the variable heavy and light chains comprise the CDRsaccording to (a); or (ii) the variable heavy chain comprises SEQ ID NO:682, the variable light chain comprises SEQ ID NO: 702, and the variableheavy and light chains comprise the CDRs according to (b); or (iii) thevariable heavy chain comprises SEQ ID NO: 722, the variable light chaincomprises SEQ ID NO: 742, and the variable heavy and light chainscomprise the CDRs according to (c); or (iv) the variable heavy chaincomprises SEQ ID NO: 762, the variable light chain comprises SEQ ID NO:782, and the variable heavy and light chains comprise the CDRs accordingto (d); or (v) the variable heavy chain comprises SEQ ID NO: 842, thevariable light chain comprises SEQ ID NO: 862, and the variable heavyand light chains comprise the CDRs according to (e).
 5. The antibody orantigen-binding fragment thereof of claim 1, which is selected from thegroup consisting of chimeric, humanized, and human antibodies orantibody fragments, and scFvs, Fab fragments, Fab′ fragments, monovalentantibody fragments, and F(ab′)2 fragments.
 6. The antibody orantigen-binding fragment thereof of claim 1, which lacks N-glycosylationand/or O-glycosylation.
 7. The antibody or antigen-binding fragmentthereof of claim 1, which comprises a human constant domain.
 8. Theantibody or antigen-binding fragment thereof of claim 7, wherein saidhuman constant domain comprises an IgG1, IgG2, IgG3, or IgG4 constantdomain.
 9. The antibody or antigen-binding fragment thereof of claim 1,which comprises an Fc region that has been modified to alter at leastone of effector function, half-life, proteolysis, or glycosylation. 10.The antibody or antigen-binding fragment thereof of claim 9, wherein theFc region contains one or more mutations that alters or eliminates N-and/or O-glycosylation.
 11. The antibody or antigen-binding fragmentthereof of claim 1, wherein the antibody or antigen-binding fragmentthereof is humanized.
 12. The antibody or antigen-binding fragmentthereof of claim 1, which when administered to a human subject inhibitsor neutralizes at least one biological effect elicited by PCSK9.
 13. Theantibody or antigen-binding fragment thereof of claim 1, which, whenadministered to a human subject, reduces serum cholesterol and/orinhibits the binding of PCSK9 to LDLR.
 14. The antibody orantigen-binding fragment thereof of claim 1, which binds to human PCSK9with a K_(D) that is less than about 100 nM.
 15. The antibody orantigen-binding fragment thereof of claim 1, wherein the antibody orantigen-binding fragment thereof is attached to one or more detectablemoieties.
 16. The antibody or antigen-binding fragment thereof of claim15, wherein the detectable moiety comprises a fluorescent dye, enzyme,substrate, bioluminescent material, radioactive material,chemiluminescent moiety, or mixtures thereof.
 17. A pharmaceuticalcomposition comprising at least one antibody or antigen-binding fragmentthereof according to claim 1 and a pharmaceutically acceptable diluent,carrier, solubilizer, emulsifier, preservative, or mixture thereof. 18.The pharmaceutical composition of claim 17, further comprising anotheractive agent.
 19. The pharmaceutical composition of claim 17, whereinthe other active agent is selected from the group consisting of thefollowing: statins, ACE inhibitors, angiotensin II receptor blockers(ARBs), antiarrhythmics, antiplatelet drugs, aspirin, beta blockers,amiodarone, digoxin, aspirin, anti-clotting agents, digoxin, diuretics,heart failure drugs, vasodilators, blood thinners, anti-cholesteroldrugs, cholestyramine, gemfibrozil, omega-3 polyunsaturated fatty acids,pantethine, anti-hypertensives, antidiabetogenic drugs, meglitinides,sulfonylurea, and thiazolidinediones.
 20. The pharmaceutical compositionof claim 19, which is lyophilized, stabilized and/or or formulated foradministration by injection.